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Chymotrypsin Sequencing Grade 11418467001
Enzyme Commission (EC) Number: 3.4.21.1 ( BRENDA | IUBMB ) General descriptionChymotrypsin Sequencing Grade is isolated as a specific protease in pure form from bovine pancreas.
Specificity
Serine endopeptidase that specifically hydrolyzes peptide bonds at the C-terminial of Tyr, Phe, and Trp. Leu, Met, Ala, Asp, and Glu are cleaved at a lower rate. Acts also upon amides and esters of susceptible amino acids. The specificity of Chymotrypsin Sequencing Grade is tested with melittin as a substrate.ApplicationUse Chymotrypsin Sequencing Grade for the hydrolysis of proteins by chymotrypsin alone or in combination with other proteases. It is suitable for peptide mapping, fingerprinting, and sequence analysis.
Quality
Purity: Free of impurities that may interfere with the specific cleavage or separation of peptides in reversed-phase HPLC.
Preparation Note
Working concentration: The recommended amount of enzyme is 1/200 to 1/20 of the quantity of protein by weight.
Storage conditions (working solution): 2 to 8 °C
A solution in 1 mM HCl can be stored for up to one week at 2 to 8 °C.
Inhibitors: Aprotinin, DFP, PMSF, phenothiazine-N-carbonyl chloride, TPCK, ZPCK, 2-macroglobulin, 1-antitrypsin, soybean trypsin inhibitor, and chymostatin. No inhibition by APMSF.
Reconstitution
Dissolve lyophilized chymotrypsin sequencing grade in 1mM HCl. The proteins to be sequenced are dissolved in digestion buffer (100mM Tris-HCl, 10mM CaCl2, pH 7.8).Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, assay 90% form lyophilized (salt-free) specific activity 70 units/mg protein (at 25 °C, with ATEE as a substrate) mol wt Mr 25 kDa packaging pkg of 4 x 25 µg Торговая марка Roche optimum pH 7.0-9.0 shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS07,GHS08 signalword Danger hcodes H315 - H319 - H334 - H335 pcodes P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311 Target Organs Respiratory system RIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Citrate Lyase (CL) 10354074001
Enzyme Commission (EC) Number: 4.1.3.6 ( BRENDA | IUBMB ) General descriptionCitrate lyase is considered as a substrate- induced enzyme which helps in catalyzing reversible aldol cleavage of citrate to oxaloacetate and acetate. The enzyme is adaptive to anaerobic dissimilation of citrate.ApplicationIt was used in enzymatic assay to measure citrate concentration using microtiter plates.
Quality
Contaminants: <0.05% ICDH (NAD specific) and “NADH oxidase”, each
Physical form
Lyophilizate, stabilized with BSA, sucrose, MgSO4 and EDTA. Note: A solution in double-distilled water has a pH-value of approximately 7.0. The enzyme can be relyophilized.
Preparation Note
Activator: Mg2+ (Km = 3.0 mM, pH 7.4, 25 °C); other divalent metal ions (Mn2+, Fe2+, Zn2+)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form lyophilized specific activity 0.25 U/mg (At 25 °C with citrate as the substrate.) packaging pkg of 120 U Торговая марка Roche optimum pH 8.0-9.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash Узнать цену -
Colcemid 10295892001
Empirical Formula (Hill Notation): C21H25NO5 CAS Number: 477-30-5 Molecular Weight: 371.43 Beilstein/REAXYS Number: 2822892 MDL number: MFCD00075459 PubChem Substance ID: 329749009 General descriptionColcemid is also known as demecolcine. Its generic name is N-methyl-N-deacetyl-colchicine. Colcemid depolymerizes microtubules and blocks mitosis at metaphase.ApplicationColcemid inhibits the formation of mitotic spindles. It is used to increase the percentage of metaphase cells for chromosome analysis.
Biochem/physiol Actions
Often in karyotyping and cell cycle research it is desirable to increase the yield of mitotic cells in a particular phase of the cell cycle. This can be achieved in a variety of ways with the most popular being the use of a cell cycle synchronizing agent such as demecolcine. Demecolcine will arrest cells in metaphase with no remarkable effect on the biochemical events in mitotic cells or in synchronized G1 and S phase cells. White blood cells are often treated with demecolcine to arrest cells in metaphase.
Physical form
Solution (10 μg/ml), filtered through 0.2 μm pore size membrane.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility sterile; 0.2 µm filtered form solution packaging pkg of 20 mL (10 µg/ml) Торговая марка Roche application(s) blocking: suitable impurities microbial, tested solubility water: miscible shipped in wet ice storage temp. 2-8°C SMILES string CN[C@H]1CCc2cc(OC)c(OC)c(OC)c2C3=CC=C(OC)C(=O)C=C13 InChI 1S/C21H25NO5/c1-22-15-8-6-12-10-18(25-3)20(26-4)21(27-5)19(12)13-7-9-17(24-2)16(23)11-14(13)15/h7,9-11,15,22H,6,8H2,1-5H3/t15-/m0/s1 InChI key NNJPGOLRFBJNIW-HNNXBMFYSA-N
Safety InformationRIDADR NONH for all modes of transport WGK Germany nwg Flash Point F does not flash Flash Point C does not flash Узнать цену -
Collagen 11179179001
General descriptionCollagens are abundantly present in vertebrates. 41 specific genes codes for 42 polypeptide chains and produce 27 collagen types. Collagens are mainly present in the extracellular matrix. Collagen type I is abundantly present in mammals. Type IV collagen is an important structural protein of basement membranes. It is chemically and genetically different from stroma collagen types I and III and cartilage collagen type II.
Specificity
Active on most mammalian cells.ApplicationCollagen is used as a substrate for culturing a variety of cells, such as coating of culture dishes and the preparation of collagen gels.
Collagen has been used in the preparation of coverslip coating solution and collagen gel contraction assay.
Biochem/physiol Actions
Collagen provides extracellular support for multicellular animals. Collagen type I offers mechanical stability, strength and toughness to a range of tissues from tendons and ligaments, to skin, cornea, bone and dentin.
Features and Benefits
Contents
Lyophilizate, cell culture grade, 3 vials of 10mg
Specifications
Biological activity: Tested for the promotion of adherence of HUV-EC cells.
Preparation Note
Working concentration: 5 µg/cm2
5 µg/cm2 for the coating of cell culture vessels; for the preparation of collagen gels, a final concentration of 2 to 3 mg/ml is used.
Reconstitution
IFor best results, dissolve the lyophilizate in 0.2% acetic acid (v/v) solution.
For the preparation of collagen gels, the content of the bottle should be dissolved in 3.3 ml sterile 0.2% acetic acid (v/v) each. This results in a final concentration of 3 mg/ml.
For coating culture dishes the final concentration should be 1 to 2 mg/ml.
Note: For dissolving: do not stir, just pour the acetic acid onto the lyophilizate and allow it to stand for several hours until it has dissolved. To completely dissolve the product an incubation for up to a maximum of 24 hours at 15 to 25 °C may be required.
Reconstitution of the Collagen should be done at 15 to 25 °C, higher temperatures destroy the fibres and may prevent the subsequent gelation of the Collagen.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыbiological source rat tail Quality Level 100 sterility sterile form lyophilized (clear, colorless solution after reconstitution) packaging pkg of 30 mg Торговая марка Roche application(s) cell culture | mammalian: suitable UniProt accession no. P02454, shipped in wet ice storage temp. 2-8°C Gene Information rat ... Col1a1(29393)
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash Узнать цену -
Collagenase B
Enzyme Commission (EC) Number: 3.4.24.3 ( BRENDA | IUBMB )
Кат. номер 11088807001 11088831001 11088815001 General descriptionCollagenase B is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization. Collagenase from Clostridium histolyticum is a collagenolytic enzyme. It interacts with collagen-like substances and is responsible for the degradation of collagen into small peptides. It is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single-cell suspensions for the establishment of primary cell culture systems. Collagenase B is recommended when yield and viability are important.
Specificity
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.ApplicationClostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.
Collagenase B has been used for preparation of cells from mouse aortas and colorectal cancer tissue.
Specifications
Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase B has a normal to high collagenase activity and a higher than average clostripain activity (usually >10U/mg).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean), serum
Unit Definition
Collagenase from Roche is assayed in Wnsch units (1 µmol of product formed per minute at +25 °C with Wnsch substrate).
Frequently, collagenase activities are given in Mandl units (1 µmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wnsch U/mg and 250 Mandl U/mg).
Preparation Note
Activator: Ca2+
Working concentration: 0.5 to 2.5 mg/ml
Storage conditions (working solution): -15 to -25 °C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25 °C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Reconstitution
Reconstitution in any balanced salt solution (e.g., HBSS)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility non-sterile form lyophilized packaging pkg of 100 mg (11088807001), pkg of 2.5 g (11088831001), pkg of 500 mg (11088815001) Торговая марка Roche concentration 0.5-2.5 mg/mL optimum pH 6.0-8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
Collagenase D
Enzyme Commission (EC) Number: 3.4.24.3 ( BRENDA | IUBMB )
Кат. номер 11088858001 11088882001 11088866001 General descriptionCollagenase D is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.SpecificityCollagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.ApplicationCollagenase from C. histolyticum is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single cell suspensions for the establishment of primary cell culture systems. Collagenase D is recommended when functionality and integrity of cell-surface proteins are important.
Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.Features and BenefitsContents
Lyophilizate, nonsterileUnit DefinitionCollagenase from Roche is assayed in Wnsch units (1 µmol of product formed per minute at +25 °C with Wnsch substrate).
Frequently, collagenase activities are given in Mandl units (1 µmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wnsch U/mg and 250 Mandl U/mg).Physical formLyophilizate, nonsterile.Preparation NoteActivator: Ca2+
Working concentration: 0.5 - 2.5mg/ml
Storage conditions (working solution): -15 to -25°C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25°C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean)
Enzyme activity:
Collagenase activity: >0.15 U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase D has a normal to high collagenase activity and very low tryptic activity (usually <0.2 units/mg, BAEE as substrate).ReconstitutionReconstitution in any balanced salt solution (e.g., HBSS)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100 sterility non-sterile form lyophilized packaging pkg of 100 mg (11088858001), pkg of 2.5 g (11088882001), pkg of 500 mg (11088866001) Торговая марка Roche concentration (working concentration: 0.5 - 2.5 mg/ml) optimum pH 6.0-8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
Collagenase H
Enzyme Commission (EC) Number: 3.4.24.3 ( BRENDA | IUBMB )
Кат. номер 11074032001 11087789001 11074059001 General descriptionCollagenase H is an enzyme mixture prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.
Specificity
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases also degrade other proteins.ApplicationCollagenase from C. histolyticum is used for the dissociation of tissues for the establishment of primary cell cultures. Collagenase H is well suited for the isolation of hepatocytes from rat liver by the collagenase perfusion method. This enzyme preparation is also suitable for the preparation of other types of cells such as endothelial cells from large vessels, and for the isolation of adipocytes from epididymal fat pads of rats.
Physical form
Lyophilizate, nonsterile
Preparation Note
Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate).
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase H has a balanced ratio of enzyme activities, and is function tested for the isolation of rat hepatocytes (perfusion method).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean)
Activator: Ca2+
Working concentration: 0.5 to 2.5mg/ml,
Approximately 1 mg/ml for the isolation of rat hepatocytes and 2mg/ml for the isolation of adipocytes.
Storage conditions (working solution): -15 to -25°C
The reconstituted solution should be stored at -15 to -25°C for short term storage, -60°C or as described below for long-term storage.
Reconstitution
Reconstitution in any balanced salt solution (e.g., HBSS)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility non-sterile form lyophilized packaging pkg of 100 mg (11074032001), pkg of 2.5 g (11087789001), pkg of 500 mg (11074059001) Торговая марка Roche optimum pH 6.0-8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
cOmplete™ His-Tag Purification Column
Кат. номер 6781543001 6781535001 General descriptionThe most common technique in affinity purification of proteins involves engineering a sequence of 6 to 14 histidines into the N- or C-terminal (or even on an exposed loop) of the protein. Such polyhistidine stretches bind strongly to divalent metal ions such as nickel and cobalt. This effect can be used to separate proteins. Metal ions can be immobilized on a matrix using a chelator, which still allows the ion to interfere with the polyhistidine tag of the protein. When these his-tagged proteins are passed through a column containing immobilized metal ions, the proteins bind via the tag to the column. Nearly all untagged proteins pass through the column. The protein is released from the column by elution with either imidazole, which competes with the polyhistidine tag for binding to the column, or by a decrease in pH, which decreases the affinity of the tag for the resin. While this procedure is generally used for the purification of recombinant proteins with an engineered affinity tag, it can also be used for natural proteins with an inherent affinity for divalent cations.
Features and Benefits
cOmplete His-Tag Purification Columns are available in 2 sizes and prepacked with cOmplete His-Tag Purification Resin. The cOmplete His-Tag Purification Resin is an innovative high-capacity IMAC matrix (Immobilized Metal Affinity Chromatography) for convenient single-step purifications of His-tagged proteins from total lysates. Roches propriety nickel-chelate chemistry ensures extraordinary compatibility with commonly used reducing agents such as DTT, chelating metalloprotease inhibitors such as EDTA, and a wide range of buffer substances and salt conditions. The wide choice of compatible ingredients allows optimization of buffers for maximum protein stability and solubility.
cOmplete His-Tag Purification Columns are fully compatible with standard purification systems such as KTA Systems (Cytiva).
- Use the buffer conditions best suited to your protein
- Repeatedly obtain highly pure protein
- Protect your protein from toxic nickel
- Work in a safe and eco-friendly environment
Packaging- 06781535001 (5 mL column; 1 column containing 5 ml of cOmplete His-Tag Purification Resin)
- 06781543001 (5 x 1 mL columns; 5 columns containing 1 ml of cOmplete His-Tag Purification Resin)
Compatibility
Compatibility for long term storage: 20% ethanol, pH 4.0 to pH 9.0
Compatibility during chromatography: The resin is compatible with 10mM EDTA, 10mM DTT during the purification (1 hour incubation), 6M guanidinium-HCl, 8M urea, pH 2.0 to pH 14.0.
Compatibility during cleaning: 4% SDS Form cOmplete His-Tag Purification Resin filled in columns, pre-charged with Ni2+ stored in 20% ethanol.Other NotesRecommended volumetric flow rate: 5 ml column (06 781 535 001): 2.5 to 10 ml/minute
1 ml column (06 781 543 001): 0.5 to 2.0 ml/minute
The volumetric flow rate is a function of the columns cross section.
Recommended imidazole concentration for load/wash: Nonspecific binding of proteins without a His-tag is low.
Use up to 5 mM imidazole in load and/or wash buffers.
If establishing a new assay for purification of His-tag proteins with cOmplete His-Tag Purification Columns do not use imidazole. If, e.g., the purity of the His-tag protein needs to be improved following this first step, use imidazole in a final concentration of up to 5mM in a second step.
Recommended imidazole concentration for elution: Up to 500mM
Please note:
In contrast to other available resins, bound His-tagged protein typically elutes from cOmplete His-Tag Purification Columns with a lower imidazole concentration, e.g., 25 to 45mM.
Binding capacity
The binding capacity of the resin to various types of proteins may vary according to the protein characteristics such as the size of the protein.
cOmplete His-Tag Purification Columns bind with a high specificity to the polyhistidine-tagged protein. As a consequence, the binding kinetics may appear to be different when compared to conventional metal chelate matrices. Full capacity of cOmplete His-Tag Purification Columns can be achieved by allowing more time for the protein to bind to the resin by lowering the flow rate during the chromatography purification procedure.
For life science research only. Not for use in diagnostic procedures.Legal InformationcOmplete is a trademark of RocheПараметрыpackaging pkg of 5 x 1 mL (columns (06781543001)), pkg of 5 mL (column (06781535001)) Торговая марка Roche parameter 1,420 cm/hr max. flow rate capacity 40 mg/mL, resin binding capacity shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
cOmplete™ His-Tag Purification Resin
Кат. номер 5893682001 5893801001 General descriptionBead size: 45 to 165µM
Binding capacity: 40mg protein per ml bed volume of resin. The binding capacity of the resin to various types of proteins may vary according to the protein characteristics such as the size of the protein. cOmplete His-Tag Purification Columns bind with a high specificity to the polyhistidine-tagged protein. As a consequence, the binding kinetics may appear to be different when compared to conventional metal chelate matrices. Full capacity of cOmplete His-Tag Purification Columns can be achieved by allowing more time for the protein to bind to the resin by lowering the flow rate during the chromatography purification procedure.
Maximal linear flow rate: 1,420cm/hour
Recommended volumetric flow rate: The volumetric flow rate is a function of the cross section of the column. Using the following formula, a linear flow rate can be converted to a volumetric flow (mL/min.): Linear flow rate (cm/hour) x column cross sectional area (cm2)/60. The column cross sectional area is defined as x r2, whereas is the constant pi and r is the inner radius of the column.
Recommended imidazole concentration for load/wash: Nonspecific binding of proteins without a His-tag is low. Use up to 5mM imidazole in load and/or wash buffers. If establishing a new assay for purification of His-tag proteins with cOmplete His-Tag Purification Columns, do not use imidazole. If, e.g., the purity of the His-tag protein needs to be improved following this first step, use imidazole in a final concentration of up to 5mM in a second step.
Recommended imidazole concentration for elution: Up to 500 mM. Please note: Contrary to other available resins, bound His-tagged protein typically elutes from cOmplete His-Tag Purification Columns with a lower imidazole concentration, e.g., 25 to 45mM.
Compatibility for long term storage: 20% ethanol, pH 4.0 to pH 9.0
Compatibility during chromatography: The resin is compatible with 10mM EDTA, 10mM DTT during the purification (1 hour incubation), 6M guanidinium-HCl, 8M urea, pH 2.0 to pH 14.0.
Compatibility during cleaning: 4% SDS
The cOmplete His-Tag Purification Resin is an innovative high-capacity IMAC matrix (Immobilized Metal Affinity Chromatography) for the purification of histidine-tagged proteins via batch or liquid chromatography procedures from total lysates. In contrast to all currently available resins, which are primarily based on NTA or IDA chelator chemistry, this resin is the only resin on the market which tolerates buffers that contain EDTA. Because EDTA is a known inhibitor of metalloproteases–which are frequently present in any cell type–this chromatography material provides the option to increase the protection of your target protein by supplementing your buffers with EDTA. This resin therefore allows increased protection of your proteins against degradation and, at the same time, enriches the protein using the well-established affinity purification method.
In addition, the resin maintains its binding capacity whether you are using low- or high-molecular weight proteins, and its specificity allows a one-step purification. By using one of the strongest chelators available, minimal contamination of your flow-through fractions with metal ions is ensured, safeguarding your downstream applications.
The cOmplete His-Tag Purification Resin is also available as prepacked format: with 1mL or 5mL resin.ApplicationRecombinant Protein Expression
Purifying a protein of interest is often essential for determining its function, structure, or interactions, for raising specific antibodies, or preparing enzymes for practical applications. Isolation of naturally expressed proteins from their original source can be a complex process involving numerous chromatographic steps. Recombinant protein expression in dedicated host organisms can greatly simplify this task. Such expression systems generally ensure higher expression levels. Fusing the target protein to a tag also confers advantageous binding ability to an affinity matrix.
Protein Purification using Immobilized Ni2+
The most common technique in affinity purification of proteins involves engineering a sequence of 6 to 14 histidines into the N- or C-terminal (or even on an exposed loop) of the protein. Such polyhistidine stretches bind strongly to divalent metal ions such as nickel and cobalt. This effect can be used to separate proteins. Metal ions can be immobilized on a matrix using a chelator, which still allows the ion to interfere with the polyhistidine tag of the protein. When these his-tagged proteins are passed through a column containing immobilized metal ions, the proteins bind via the tag to the column. Nearly all untagged proteins pass through the column. The protein is released from the column by elution with either imidazole, which competes with the polyhistidine tag for binding to the column, or by a decrease in pH, which decreases the affinity of the tag for the resin. While this procedure is generally used for the purification of recombinant proteins with an engineered affinity tag, it can also be used for natural proteins with an inherent affinity for divalent cations.
His-Tags
Ideally, the His-tagged target protein binds much stronger to the Ni2+ chelate matrix than endogenous histidine-containing protein of the expression host. Relative binding strength depends on how many histidines can bind simultaneously to the matrix (avidity effect). Longer His-tags confer stronger binding and better separation of the target from potentially contaminating host proteins. The classic His-tag has six consecutive histidines. Tags with 10 to 14 histidines may produce a better purification. Most importantly, His-tagged proteins can be purified using Ni2+ chelate matrices under both native and denaturing conditions. Due to their hydrophilic and flexible nature, these matrices increase the solubility of the target proteins and only rarely interfere with protein function. This unique combination of features enables the His-tag to be a versatile tool for a wide range of protein purification applications.
Features and Benefits
cOmplete His-Tag Purification Resin is the only resin for purifying large amounts of protein without compromises.
- Use the buffer conditions best suited to your protein: Keep your protein comfortable and let it, not your purification resin, determine whether you use DTT, EDTA, or other buffer substances.
- Repeatedly obtain highly pure protein: Single step purification without resin recharging.
- Protect your protein from toxic nickel: Reduce protein oxidation and aggregation caused by resins that leach nickel.
- Work in a safe and eco-friendly environment: Avoid handling of toxic nickel and completely eliminate disposal costs.
Physical form
50% resin in suspension, pre-charged with Ni2+Other NotesFor life science research only. Not for use in diagnostic procedures.
EDTA-compatible resin for the purification of poly-histidine-tagged proteins.Legal InformationcOmplete is a trademark of RocheПараметрыpackaging pkg of 200 mL (05893801001 settled resin volume), pkg of 25 mL (05893682001 settled resin volume) mfr. no. Roche matrix Sepharose-CL 6B shipped in ambient storage temp. 2-8°C
Safety Information
