Cell Health Analysis Sigma-Aldrich
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Annexin V-Cy3™ Apoptosis Detection Kit
NACRES: NA.32
Кат. номер APOAC-1KT APOAC-1KT-PW Features and BenefitsDetects apoptosis earlier in the process than DNA-based assays such as TUNEL- Rapid labeling of cells. Cell staining takes only 10 minutes.
- No cell fixation or processing required, reducing the detection time and allowing the cells to be used for further study.
- 6-Carboxyfluorescein diacetate secondary dye is included with the kit to differentiate apoptotic cells from viable and necrotic cells.
PrincipleSigma′s Annexin V-Cy3 kit allows detection of annexin V bound to apoptotic cells by fluorescence microscopy. The Annexin V-Cy3 kit uses the dye Cy3.18 as the fluorochrome conjugated with annexin V. By microscopy, Cy3.18 fluoresces more brightly than the FITC conjugate. The kit includes the non-fluorescent compound 6-carboxyfluorescein diacetate (6-CFDA), which enters the cell and is hydrolyzed by the esterases present in living cells to the fluorescent compound 6-carboxyfluorescein, indicating that the cells are viable. This combination allows the differentiation among early apoptotic cells (annexin V positive, 6-CFDA positive), necrotic cells (annexin V positive, 6-CFDA negative), and viable cells (annexin V negative, 6-CFDA positive).Legal InformationCy3 is a trademark of CytivaПараметрыusage sufficient for 200 tests Quality Level 200 packaging pkg of 1 kit application(s) cell analysis: suitable, detection: suitable, flow cytometry: suitable detection method fluorometric shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 3 Flash Point(F) Not applicable Flash Point(C) Not applicable Узнать цену -
Annexin V-FITC Apoptosis Detection Kit
Кат. номер APOAF-20TST-PW APOAF-50TST-PW APOAF-50TST APOAF-20TST APOAF-60TST General descriptionAnnexin V-FITC kit allows fluorescent detection of annexin V bound to apoptotic cells and quantitative determination by flow cytometry. The AnnexinV-FITC kit uses annexin V conjugated with fluorescein isothiocyante (FITC) to label phosphatidylserine sites on the membrane surface. The kit includes propidium iodide (PI) to label the cellular DNA in necrotic cells where the cell membrane has been totally compromised. This combination allows the differentiation among early apoptotic cells (annexin V positive, PI negative), necrotic cells (annexin V positive, PI positive), and viable cells (annexin V negative, PI negative).ApplicationAnnexin V-FITC Apoptosis Detection Kit was used:- in staining of LNCaP prostate cancer cells for measuring the G. lucidum extracts activity during the treatment of prostate cancer.
- for tumor cell labelling to study the inhibitory activity of DBP-maf (Vitamin D binding protein-macrophage activating factor) on prostate tumor cells.
- for indirect measurement of flippase activity.
Features and BenefitsDetects apoptosis earlier in the process than DNA-based assays such as TUNEL.- Rapid labeling of cells. Cell staining takes only 10 minutes.
- No cell fixation or processing required, reducing the detection time and allowing the cells to be used for further study.
- Propidium iodide secondary dye is included with the kit to differentiate apoptotic cells from viable and necrotic cells.
Other NotesAllow all components to reach room temperature before use.Параметрыusage (20 tests) Quality Level 300 packaging pkg of 1 kit application(s) cell analysis: suitable, detection: suitable, flow cytometry: suitable detection method fluorometric shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
BrdU Cell Proliferation Assay
Кат. номер QIA58-200TEST QIA58-1000TEST QIA58-1000TESTCN QIA58-200TESTCN General descriptionBrdU Cell Proliferation Assay is a non-isotopic immunoassay for quantification of BrdU incorporation into newly synthesized DNA of actively proliferating cells. It is sensitive, rapid, and easy to perform.
This proliferation assay is a non-isotopic immunoassay for quantification of BrdU incorporation into newly synthesized DNA of actively proliferating cells.
A non-isotopic enzyme immunoassay for the quantification of cell proliferation.Evaluation of cell cycle progression is essential for investigations in many scientific fields. Measurement of [3H] thymidine incorporation as cells enter S phase has long been the traditional method for the detection of cell proliferation. Subsequent quantification of [3H] thymidine is performed by scintillation counting or autoradiography. This technology is slow, labor intensive and has several limitations including the handling and disposal of radioisotopes and the necessity of expensive equipment. A well-established alternative to [3H] thymidine uptake has been demonstrated by numerous investigators. In these methods bromodeoxyuridine (BrdU), a thymidine analog replaces [3H] thymidine. BrdU is incorporated, into newly synthesized DNA strands of actively proliferating cells. Following partial denaturation of double stranded DNA, BrdU is detected immunochemically allowing the assessment of the population of cells, which are actively synthesizing DNA. The Calbiochem® BrdU Cell Proliferation Assay involves incorporation of BrdU into cells cultured in plates and BrdU immunolabeling using the cell layer as the solid phase. The resultant assay is sensitive, rapid, easy to perform and applicable to high sample throughput. In addition to evaluation of cell proliferation, information such as cell number, morphology and analysis of cellular antigens can be obtained from a single culture.
Note: 1 T = 1 test.Packaging200, 1000 test in Fibre caseComponentsBrdU Label, Fixative/Denaturing Solution, BrdU Antibody, Antibody Diluent, Peroxidase Goat Anti-Mouse IgG, Conjugate Diluent, Substrate, Plate Wash Concentrate, Stop Solution, and a user protocol.WarningToxicity: Multiple Toxicity Values, refer to MSDS (O)SpecificationsAssay Time: 3 hPrincipleThe Calbiochem® BrdU Cell Proliferation Assay is a non-isotopic immunoassay for the quantitation of bromodeoxyuridine incorporation into newly synthesized DNA of actively proliferating cells.Storage and StabilityUpon arrival store the entire contents of the kit at -20°C, in a non-frostfree freezer.
Prior to use:
1. Remove the Fixative/Denaturing Solution and place at room temperature for at least 4 h prior to use. Precipitates that may occur while cold should go back into solution.
2. Reconstitute the Peroxidase Goat Anti-Mouse IgG with the appropriate volume of 1X PBS.
• For the 200 test kit, reconstitute Peroxidase Goat Anti-Mouse IgG in 125 µl of 1X PBS. Use Conjugate Diluent for any further lot-specific dilution.
• For the 1000 test kit, reconstitute Peroxidase Goat Anti-Mouse IgG in 250 µl of 1X PBS. Use Conjugate Diluent for any further lot-specific dilution.
Allow solution to stand at room temperature for 10 min. (or until solution is clear). Divide into small aliquots; aliquots not to be used that same day should be stored at -20°C in a non-frostfree freezer.
Once the kit components have been thawed for use:
• The BrdU label and 100X Anti-BrdU Antibody should be divided into small aliquots and stored at -20°C in a non-frostfree freezer with the Peroxidase Goat Anti-Mouse IgG; avoid multiple freeze/thaw cycles.
• All the other kit components (Substrate, Antibody Diluent, Conjugate Diluent, 20X Plate Wash Concentrate, Fixative/Denaturing Solution, Stop Solution) should be stored at 4°C.Other NotesHardonk, M.J. and Harms, G. 1990. Acta histochemica, Suppl.39, 99.
Muir, D., et al. 1990. Analytical Biochemistry185, 377.
Lanier, T. L., et al. 1989. Carcinogenesis10, 1341.
Magaud, J.-P., et al. 1988. J. Immunol. Methods106, 95.
Collins, S.J. 1987. Blood70, 1233.
Porstmann, T., et al. 1985. J. Immunol. Methods82, 169.
Raza, A., et al. 1985. Cytometry6, 633.
Morstyn, G., et al. 1983. J. Clin. Invest.72, 1844.
Gratzner, H.G. 1982. Science218, 474.
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.Legal InformationCALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, GermanyПараметрыstorage conditions -20C Quality Level 100 usage sufficient for 1000 tests, sufficient for 200 tests packaging pkg of 1 96-well plate(s) Торговая марка Calbiochem® storage condition OK to freeze, avoid repeated freeze/thaw cycles input sample type intact cells
sample type intact cellsdetection method colorimetric shipped in wet ice storage temp. 20°C Узнать цену -
Cell Growth Determination Kit, MTT based CGD1-1KT
NACRES: NA.75 ApplicationFor spectrophotometric determination of viable cell number.PackagingEach kit includes 5 vials each containing 1 mL of 5 mg/ml MTT in RPMI (without phenol red) and 50 mL of MTT solvent (0.1 N HCl in anhydrous isopropanol). Volumes of kit components have been optimized for cultures grown in multi-well plates. MTT solutions is added at 10% of total culture volume, thus the number of tests per kit is dependent upon culture vessel and/or volume.Параметрыpackaging pkg of 1 kit Quality Level 200 application(s) UV/Vis spectroscopy: suitable, cell analysis: suitable, detection: suitable, protein quantification: suitable Amax 570 nm detection method colorimetric shipped in dry ice storage temp. 20°C
Safety InformationPictograms GHS02,GHS07 Signal Word Danger Hazard Statements H225 - H315 - H319 - H335 - H336 Precautionary Statements P210 - P261 - P305 + P351 + P338 RIDADR UN 3316 9 Flash Point(F) 53.6 °F - closed cup Flash Point(C) 12 °C - closed cup Узнать цену -
Flow Cytometry Kit for Apoptosis APOBRDU-1KT
NACRES: NA.32 ApplicationThe greater incorporation of Br-dUTP results in a stronger signal by flow cytometry when detected using a fluorescein-labeled anti-BrdU antibody.Propidium iodide/RNase A solution is included in the kit to counterstain the total DNA. APOBRDU has been used in flow cytometry assay in human non–small cell lung cancer. APOBRDU has been used in cell cycle and apoptosis assay in HeLa cells, and in tunel assay human neuroblastoma cell lines.Features and BenefitsGreater incorporation of Br-dUTP, resulting in improved detection than found by using biotin- or digoxigenin-conjugated dUTP or by using fluorochrome (fluorescein or BODIPY)-conjugated deoxynucleotides.PackagingThe kit is shipped in two parts, both wet ice. Upon receipt, store APO-PART1 at −20 °C and APO-PART2 at 2-4 °CPrincipleBr-dUTP is incorporated more readily into the DNA fragments than deoxyuridine triphosphate labeled with larger dyes such as FITC, biotin or digoxigenin. One of the biological characteristics that defines apoptosis is the degradation of genomic DNA into fragments of 180-200 bp, commonly called "DNA laddering". The fragmentation creates a large number of 3′-hydroxyl sites at the DNA breaks. This property is used in the APO-BRDU kit to identify apoptotic cells by labeling the 3′-hydroxyl sites with bromodeoxyuridine triphosphate (Br-dUTP). Br-dUTP is enzymatically attached to the 3-hydroxyl sites of double- or single-stranded DNA by terminal transferase (TdT). Non-apoptotic cells do not incorporate Br-dUTP due to the lack of available 3-hydroxyl sites.Параметрыusage sufficient for 60 cell suspensions packaging pkg of 1 kit application(s) cell analysis: suitable, detection: suitable, flow cytometry: suitable detection method fluorometric shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS02,GHS07,GHS08 signalword Danger hcodes H225 - H319 - H350 pcodes P201 - P202 - P210 - P233 - P305 + P351 + P338 - P308 + P313 RIDADR UN 3316 9 WGK Germany WGK 3 Flash Point F 55.4 °F Flash Point C 13 °C Узнать цену -
In Vitro Toxicology Assay Kit, Lactic Dehydrogenase based TOX7-1KT
NACRES: NA.84 General descriptionTox7 is a member of Sigmas CytoToxicity Kit series. It is based on the measurement of lactate dehydrogenase (LDH) release from cells as a measure of cell membrane damage and cytotoxicity. This technique has been utilized as an alternative to 51Cr release for cell mediated cytotoxicity assays, as well as conventional cytotoxicity resulting from interaction of a test material with the cell.ApplicationIn Vitro Toxicology Assay Kit, Lactic Dehydrogenase based has been used:- to detect lactate dehydrogenase release in human umbilical vein endothelial cells (HUVECs) and human cardiac myocytes (HCMs) for lactate dehydrogenase (LDH) assay
- to determine the activity of lactate dehydrogenase leakage of MDA-MB-231 cells
- to examine the activity of LDH in RAW 264.7 cells , in LDH cytotoxicity assay in human cell lines
- for spectrophotometric measurement of viable cells. Absorbance of converted dye is measured at a wavelength of 490 nm.
Biochem/physiol ActionsThe lactate dehydrogenase assay is a means of measuring either the number of cells via total cytoplasmic lactate dehydrogenase (LDH) or membrane integrity as a function of the amount of cytoplasmic LDH released into the medium. This method is simple, accurate and yields reproducible results. The assay is based on the reduction of NAD by LDH. The resulting reduced NAD (NADH) is utilized in the stoichiometric conversion of a tetrazolium dye to a soluble, colored formazan derivative.
.ПараметрыQuality Level 200 form liquid usage 1 kit sufficient for 500 tests packaging pkg of 1 kit storage condition dry at room temperature application(s) cell analysis: suitable, detection: suitable, protein quantification: suitable max 490 nm detection method colorimetric storage temp. 20°C
Safety InformationHazard Statements H412 Precautionary Statements P273 - P501 RIDADR NONH for all modes of transport WGK Germany WGK 3 Узнать цену -
In Vitro Toxicology Assay Kit, MTT based TOX1-1KT
NACRES: NA.84 General descriptionThis kit is designed for determining cell number/ cell viability spectrophotometrically as a function of mitochondrial activity in living cells. The MTT method is simple, accurate and yields reproducible results. The key component is (3- [4,5- dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) or MTT. Solutions of MTT, dissolved in medium or balanced salt solutions without phenol red, are yellowish in color. Mitochondrial dehydrogenases of viable cells cleave the tetrazolium ring, yielding purple formazan crystals which are insoluble in aqueous solutions. Absorbance of converted dye is measured at a wavelength of 570nm.ApplicationIn Vitro Toxicology Assay Kit, MTT based has been used to perform an (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide) MTT assay to measure the cytotoxicity of gold nanoparticles (AuNP). It has also been used to examine cell viability in SH-SY5Y cultures in response to 6-hydroxydopamine (6-OHDA) treatment.Biochem/physiol ActionsConversion of MTT to a water-insoluble colored formazan derivative which is then solubilized in acidic isopropanol.ПараметрыQuality Level 200 usage 1 kit sufficient for 1,000 tests packaging pkg of 1 kit storage condition dry at room temperature application(s) cell analysis: suitable, detection: suitable max 570 nm detection method colorimetric storage temp. 2-8°C
Safety InformationPictograms GHS02,GHS05,GHS07,GHS08 Signal Word Danger Hazard Statements H225 - H314 - H335 - H336 - H341 - H412 Precautionary Statements P210 - P261 - P273 - P280 - P305 + P351 + P338 - P310 RIDADR UN 3316 9 Flash Point(F) 53.6 °F Flash Point(C) 12 °C Узнать цену -
In Vitro Toxicology Assay Kit, Neutral Red based TOX4-1KT
NACRES: NA.84 General descriptionFor spectrophotometric measurement of viable cells. Absorbance of converted dye is measured at a wavelength of 540 nm.Biochem/physiol ActionsNeutral red is taken up by viable cells and stored in the lysosomes. The dye is extracted and the uptake is quantitated by spectroscopy.Параметрыform liquid Quality Level 200 usage 1 kit sufficient for 1,000 tests packaging pkg of 1 kit storage condition dry at room temperature application(s) cell analysis: suitable, detection: suitable max 540 nm detection method colorimetric storage temp. 2-8°C
Safety InformationPictograms GHS02,GHS07,GHS08 Signal Word Danger Hazard Statements H226 - H317 - H319 - H350 Precautionary Statements P202 - P210 - P280 - P303 + P361 + P353 - P305 + P351 + P338 - P308 + P313 RIDADR UN 3316 9 Flash Point(F) 73.4 °F Flash Point(C) 23 °C Узнать цену -
In Vitro Toxicology Assay Kit, Resazurin based TOX8-1KT
EC Number: 263-718-5 NACRES: NA.84 ApplicationFor spectrophotometric measurement of metabolic activity of living cells. Absorbance of converted dye is measured at a wavelength of 600 nm.Biochem/physiol ActionsBioreduction of the dye reduces the amount of its oxidized form (blue) and concomitantly increases the fluorescent intermediate (red).ПараметрыQuality Level 200 form liquid usage 1 kit sufficient for 2,000 tests packaging pkg of 1 kit storage condition dry at room temperature application(s) cell analysis: suitable, detection: suitable, protein quantification: suitable detection method colorimetric, fluorometric storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 3 Flash Point(F) Not applicable Flash Point(C) Not applicable Узнать цену -
LookOut® Mycoplasma Elimination Kit
NACRES: NA.75
Кат. номер MP0030-1KT MP0030-1KT-PW General descriptionThe kit is comprised of a combination of biological agents that reliably and completely eliminate mycoplasma contamination. The initial treatment of this eradication procedure is adequate for mycoplasma elimination in most applications. The second step suppresses and inactivates any remaining mycoplasma using a follow-up antibiotic treatment.ApplicationThe LookOut® Mycoplasma Elimination Kit has been developed to quickly and efficiently eliminate mycoplasma contamination from cell cultures.Features and Benefits- Minimal cytotoxic properties of the reagents
- Highly effective
- Retention of normal cell characteristics
SuitabilityThe LookOut® Mycoplasma Elimination Kit is suitable for the elimination of Mollicutes and related organisms (Mycoplasma, Acholeplasma, Spiroplasma, and Entomoplasma) in cell and virus cultures. It is not effective for treating mycoplasma contamination in Chlamydia cultures.Legal InformationLookOut is a registered trademark of Sigma-Aldrich Co. LLCПараметрыQuality Level 200 usage 1 kit sufficient for 5 mL mycoplasma elimination packaging pkg of 1 kit storage condition protect from light application(s) PCR: suitable, detection: suitable, microbiology: suitable storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
LookOut® Mycoplasma qPCR Detection Kit MP0040A-1KT
EC Number: 200-664-3 General descriptionThe LookOut Mycoplasma PCR Detection Kit utilizes the polymerase chain reaction (PCR), which is established as the method of choice for highest sensitivity in the detection of Mycoplasma and Acholeplasma contamination in cell cultures and other cell culture derived biologicals.
The primer/probe system detects the highly conserved 23S rRNA operon coding region of the mycoplasma genome. The kit is highly specific and does not detect eukaryotic DNA. The detection spectrum includes most mycoplasma species identified as cell culture contaminants (see Table 2 in the technical bulletin). The kit contains the nucleotide dUTP instead of dTTP and is, therefore, suitable for UNG pretreatment.
This kit already contains Taq polymerase in the Mycoplasma Mix.ApplicationqPCR Probe Kit for high quality and reliable quantitative detection of mycoplasma DNA in research, industrial application and product testing. Usable for direct testing of cell cultures and biologicals in combination with cell culture enrichment.Legal InformationLookOut is a registered trademark of Sigma-Aldrich Co. LLCПараметрыQuality Level 200 usage sufficient for 25 reactions quality Not suitable for clinical diagnostic use. packaging pkg of 1 kit storage condition dry at room temperature application(s) PCR: suitable, detection: suitable, microbiology: suitable storage temp. 2-8°C
Safety InformationFlash Point(F) 188.6 °F Flash Point(C) 87 °C Узнать цену -
MTT Cell Growth Assay Kit CT02
eCl@ss: 32161000 NACRES: NA.32 General descriptionMTT is a pale yellow substrate that is cleaved by living cells to yield a dark blue formazan product. This process requires active mitochondria, and even freshly dead cells do not cleave significant amounts of MTT. The colorimetric assay described below can be used for either proliferation or complement-mediated cytotoxicity assays.ApplicationProcedure:
1) Carry out a lymphokine, mitogen, or complement-mediated cytotoxicity assay using standard methods, in 96-well flat-bottomed tissue culture plates of good optical quality (e.g. Falcon). The final volume of tissue culture medium in each well should be 0.1 mL, and the medium (e.g. RPMI or DMEM) may contain up to 10% Fetal Bovine Serum.
2) At the end of the assay add 0.01 mL AB Solution (MTT) to each well. Mix by tapping gently on the side of the tray.
3) Incubate at 37° C for cleavage of MTT to occur. Optimal times may vary according to the assay, but four hours is suitable for most purposes. At the end of this time, the MTT formazan produced in wells containing live cells will appear as black, fuzzy crystals on the bottom of the well.
4) Add 0.1 mL isopropanol with 0.04 N HCl to each well. Mix thoroughly by repeated pipetting with a multichannel pipettor. The HCl converts the phenol red in tissue culture medium to a yellow color that does not interfere with MTT formazan measurement. The isopropanol dissolves the formazan to give a homogeneous blue solution suitable for absorbance measurement.
5) Within an hour, measure the absorbance on an ELISA plate reader with a test wavelength of 570 nm and a reference wavelength of 630 nm. After several hours at room temperature, serum proteins may begin to precipitate due to the acid/alcohol. Chilling the plates will hasten the precipitation. If the plates must be stored before measuring, keep at 4° C before adding the acid/alcohol, then warm to room temperature and add acid/alcohol just before reading.
Results:
The MTT assay will normally detect 200 to 50,000 cells of a typical cell line, although 1,000 to 50,000 is the useful range. This number may vary for other cell types. Cytotoxic assays should be set up so that the control, unlysed cells give a signal of 0.2 to 0.4, and proliferation assays should yield a similar value at plateau concentrations. This corresponds to about 20-50,000 cells per well with a typical cell line.
Absorbance is directly proportional to the number of cells; actual cells do not absorb significantly, even up to concentrations of 1 x 106 cells/mL.ComponentsReagent A: MTT, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), 50 mg/vial.
Solution B: PBS pH 7.4, 15 mLStorage and StabilityMaintain at 2-8°C for up to six months. The Reagent A / Solution B mixture is stable at 2-8°C for up to two weeks.
Reagent Preparation:
Add 10 mL Solution B to one vial of Reagent A. Mix well, sterile filter and keep in the dark at 4° C until used. Note: It may take overnight to dissolve. Do not heat solution. When absolutely required to dissolve crystals, adjust pH with 1-2 drops of HCl. The AB mixture is stable for several weeks under these conditions.Legal InformationCHEMICON is a registered trademark of Merck KGaA, Darmstadt, GermanyDisclaimerUnless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.ПараметрыQuality Level 100 Торговая марка Chemicon® application(s) activity assay: suitable, cell analysis: suitable (cell based assay) shipped in wet ice Узнать цену -
MTT Cell Growth Assay Kit CT01
eCl@ss: 32161000 NACRES: NA.32 General descriptionMTT is a pale yellow substrate that is cleaved by living cells to yield a dark blue formazan product. This process requires active mitochondria, and even freshly dead cells do not cleave significant amounts of MTT. The colorimetric assay described below can be used for either proliferation or complement-mediated cytotoxicity assays.ApplicationProcedure:
1. Carry out a lymphokine, mitogen, or complement-mediated cytotoxicity assay using standard methods, in 96-well flat-bottomed tissue culture plates of good optical quality (e.g. Falcon). The final volume of tissue culture medium in each well should be 0.1 mL, and the medium (e.g. RPMI or DMEM) may contain up to 10% Fetal Bovine Serum.
2. At the end of the assay add 0.01 mL AB Solution (MTT) to each well. Mix by tapping gently on the side of the tray.
3. Incubate at 37°C for cleavage of MTT to occur. Optimal times may vary according to the assay, but four hours is suitable for most purposes. At the end of this time, the MTT formazan produced in wells containing live cells will appear as black, fuzzy crystals on the bottom of the well.
4. Add 0.1 mL isopropanol with 0.04 N HCl to each well. Mix thoroughly by repeated pipetting with a multichannel pipettor. The HCl converts the phenol red in tissue culture medium to a yellow color that does not interfere with MTT formazan measurement. The isopropanol dissolves the formazan to give a homogeneous blue solution suitable for absorbance measurement.
5. Within an hour, measure the absorbance on an ELISA plate reader with a test wavelength of 570 nm and a reference wavelength of 630 nm. After several hours at room temperature, serum proteins may begin to precipitate due to the acid/alcohol. Chilling the plates will hasten the precipitation. If the plates must be stored before measuring, keep at 4° C before adding the acid/alcohol, then warm to room temperature and add acid/alcohol just before reading.
Results:
The MTT assay will normally detect 200 to 50,000 cells of a typical cell line, although 1,000 to 50,000 is the useful range. This number may vary for other cell types. Cytotoxic assays should be set up so that the control, unlysed cells give a signal of 0.2 to 0.4, and proliferation assays should yield a similar value at plateau concentrations. This corresponds to about 20-50,000 cells per well with a typical cell line.
Absorbance is directly proportional to the number of cells; actual cells do not absorb significantly, even up to concentrations of 1 x 106 cells/mL.
MTT Cell Growth Assay is a colorimetric assay that can be used for either proliferation or complement-mediated cytotoxicity assays.ComponentsReagent A: MTT, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), 50 mg/vial.
Solution B: PBS pH 7.4, 60 mLStorage and StabilityMaintain at 2-8°C for up to six months. The Reagent A / Solution B mixture is stable at 2-8°C for up to two weeks.
Reagent Preparation:
For every 1,000 assays to be performed, add 10 mL Solution B to one vial of Reagent A. Mix well, sterile filter and keep in the dark at 4° C until used. Note: It may take overnight to dissolve. Do not heat solution. When absolutely required to dissolve crystals, adjust pH with 1-2 drops of HCl. The AB mixture is stable for several weeks under these conditions.Legal InformationCHEMICON is a registered trademark of Merck KGaA, Darmstadt, GermanyDisclaimerUnless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.ПараметрыQuality Level 100 Торговая марка Chemicon® application(s) activity assay: suitable, cell analysis: suitable (cell based assay) shipped in wet ice Узнать цену -
Rapid Cell Proliferation Kit
Кат. номер QIA127-1KIT QIA127-1KITCN General descriptionAssay measures the increased activity of cellular mitochondrial dehydrogenases that can cleave the tetrazolium dye WST-1 to formazan. The formazan formation is then quantified by measuring the change in absorbance at 450 nm in a microplate reader. The activity of mitochondrial dehydrogenases is proportional to cell number. No washing, harvesting, or solubilization steps are required.Packaging1 kit in Glass bottleFeatures and BenefitsSensitivity: 1000 cells (adherent); 2500 cells (nonadherent)ComponentsWST-1, 1-Methoxy PMS, and a user protocol.WarningToxicity: Multiple Toxicity Values, refer to MSDS (O)SpecificationsAssay Time: 0.5 - 4 hPrincipleThe Calbiochem Rapid Cell Proliferation Kit is designed to determine the number of viable cells in proliferation, or cytotoxicity studies, based on the cleavage of the tetrazolium salt WST-1 by mitochondrial dehydrogenases in viable cells. The assay can be used to measure cell proliferation in response to growth factors or cytokines, or to assess cytotoxic compounds.Preparation Note• WST-1 Labeling Mixture: Thaw the EMS solution at room temperature or in a 37°C water bath. Add the entire contents of this solution to the bottle containing the WST-1 reagent. The mixed solution can be stored at 4°C for several weeks. For long-term storage (several months) aliquot and store at -20°C. Protect from light.Other NotesIshiyama, M., et al. 1995. In Vitro Toxicology8, 187.
Liu, S.O., et al 1995. Nat. Med.1, 267.
Ishiyama, M., et al. 1993. Chem. Pharm. Bull.41, 1118.Legal InformationCALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, GermanyПараметрыstorage conditions -20C Quality Level 100, usage sufficient for 500 tests packaging pkg of 1 96-well plate(s) Торговая марка Calbiochem® storage condition OK to freeze, avoid repeated freeze/thaw cycles, protect from light input sample type cultured cells test parameters sample: 1000 - 50,000 cells detection method colorimetric shipped in wet ice storage temp. 20°C Узнать цену -
Senescence Cells Histochemical Staining Kit
NACRES: NA.32
Кат. номер CS0030-1KT CS0030-1KT-PW General descriptionThe Senescence Cells Histochemical Staining Kit contains all the reagents required for identifying senescent cells using an assay based on a histochemical stain for b-galactosidase activity at pH 6. b-galactosidase activity is detectable in senescent cells, but not in quiescent, immortal, or tumor cells.ApplicationFor use in histochemical staining of b-galactosidase actifvity.Biochem/physiol ActionsReplicative senescence is a growth-arrest state associated with loss of division potential, changes in cell morphology, shape and physical appearance, and the pattern of gene expression in cells.Legal InformationManufactured under license to US Patent Nos. 5,491,069 and 5,795,728.ПараметрыQuality Level 200 usage sufficient for 100 tests packaging pkg of 1 kit storage condition dry at room temperature detection method colorimetric shipped in dry ice storage temp. −20°C
Safety InformationPictograms GHS05,GHS06,GHS08 Signal Word Danger Hazard Statements H302 + H312 - H314 - H317 - H331 - H334 - H335 - H341 - H350 - H412 Precautionary Statements P273 - P280 - P301 + P312 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 Target Organs Respiratory system RIDADR UN 3316 9 Flash Point(F) Not applicable Flash Point(C) Not applicable Узнать цену -
Senescence Detection Kit
Кат. номер QIA117-1KIT QIA117-1KITCN General descriptionUseful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. Higher levels of β-Galactosidase is considered as a good marker for senescent cells. The mechanism of this is believed to be due to the increased lysosomal content of senescent cells, resulting in an increase of the lysosomal enzyme β-galactosidase. Cells can be observed using microscope.
Senescence is thought to be a tumor suppressive mechanism and an underlying cause of aging. It represents an arrested state in which the cells remain viable, but not stimulated to divide by serum or by passage in culture. Senescent cells display an increase in cell size, pH-dependent expression of β-galactosidase (β-Gal) activity, and altered patterns of gene expression.Packaging1 kit in Glass bottleComponentsFixative Solution, X-Gal, Staining Solution, Staining Supplement, and a user protocol.WarningToxicity: Multiple Toxicity Values, refer to MSDS (O)SpecificationsAssay Time: 30 min. plus overnight incubationPrincipleThe Calbiochem® Senescence Detection Kit is useful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. The mechanism of this is believed to be the increased lysosomal content of senescent cells resulting in an increase of the lysosomal enzyme -galactosidase.Preparation Note• PBS: Prepare a 1X PBS solution (not provided). Use at least 3 ml per well.• X-gal: Dissolve 20 mg X-gal in 1 ml of DMF (N-N-dimethylformamide, not provided) to prepare a 20X stock solution. Aliquot and freeze (-20°) the unused X-gal solution. This stock solution is stable for up to 1 month at -20°C (protect from light). Always store X-gal in a polypropylene or glass container. Do not use polystyrene.• Staining Solution Mix: While cells are incubating in Fixative Solution (step 2 below), prepare Staining Solution Mix in a polypropylene plastic tube by adding 470 µl of Staining Solution, 5 µl Staining Supplement, and 25 µl of 20 mg/ml X-gal in DMF.Storage and StabilityUpon arrival store the entire contents of the kit at -20°C.Other NotesKurz, D.J., et al. 2000. J. Cell Sci.113, 3613.
Dimri, G.P., et al. 1995. Proc. Natl. Acad. Sci. USA92, 9363.Legal InformationCALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, GermanyПараметрыstorage conditions -20C usage sufficient for 250 tests Quality Level 100, packaging pkg of 1 12-well plate(s) Торговая марка Calbiochem® storage condition OK to freeze, protect from light input sample type intact cells detection method colorimetric shipped in wet ice storage temp. 20°C Узнать цену
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