Recombinant Trypsin
Trypsin specifically cleaves C-terminal peptide bonds of lysine and arginine,which can degrade intercellular binding proteins. The amino acid sequence of the recombinant trypsin produced by Hyasen Biotech is the same as that of porcine pancreas-derived trypsin, and is produced by recombinant Escherichia coli expression. It can replace traditional extraction of trypsin for cell digestion in vaccines,stem cells, immune cell therapy, drug screening, antibodies and other fields. Aprotinin, soybean trypsin inhibitor, etc. can distinctly inhibit trypsin activity.
Recombinant trypsin has the same properties as naturally extracted trypsin. This product is manufactured in a GMP grade workshop. No animal origin raw materials are used in the fermentation, purification and final ingredients. At the same time, the production and quality control processes are validated, resulting in a high level of consistency between batches.
Product features
|
CAS |
9002-07-7 |
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EC |
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|
Character |
White or off white crystalline power |
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Molecular weight |
24kDa |
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Isoelectric point |
8.26 |
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Purity (RP-HPLC) |
β-trypsin≥70.0%; α-Trypsin≤20.0% |
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Specific Activity |
≥3800.0U/mg protein (BAEE) |
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Recombinant trypsin activity |
≥2500.0U/mg power (BAEE) |
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Identification |
Purple |
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Optimal pH |
8.0 |
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Bioburden |
≤300CFU/g |
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E. Coli host cell protein residue |
≤0.01% |
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E. Coli host cell DNA residue |
≤10.0000ng/mg |
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Endotoxin content (LAL-Test) |
≤20.0EU/mg |
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Protein content |
95.0%±10.0% |
Storage conditions
Stability of storage: Recombinant trypsin lyophilized power should be stored under 2-8℃ . It is stable within 24 months. After dissolved with 1mM HCl or 50mM HAC, it should be stored under -25 ～ - 15℃ . It is no activity loss after 10 times repeated freezing and thawing.
Unit definition
25℃ , pH 7.6, 3.2ml reaction solution(1cm light path), one trypsin unit (U) was defined as an increase of 0.003 in the absorption value at 253nm by enzymatic hydrolysis of BAEE per minute.
Quality control
Activity: Determination the activity of trypsin by Pharmacopoeia of the People’s Republic of China Volume 4 (2020 Edition).
.E. Coli host cell protein residue: ELISA Kit. Pharmacopoeia of the People’s Republic of China Volume 4 (2020 Edition) -E. Coli host Cell protein residue (3412).
.Protein purity: Pharmacopoeia of the People’s Republic of China Volume 4 (2020 Edition)-General Rules for HPLC Method (0512).
. Bioburden: Pharmacopoeia of the People’s Republic of China Volume 4 (2020 Edition)-General Rules for Sterility Test (1101). • E. Coli host cell DNA residue: Pharmacopoeia of the People’s Republic of China Volume 4 (2020 Edition) Host Cell DNA residues (3407). . Endotoxin content: Pharmacopoeia of the People’s Republic of China Volume 4 (2020 Edition)-General Rules for LAL-Test (1143).
Product instruction
1.Solution preparation: Take an appropriate amount of trypsin according to the protein content (calculated by power activity and protein activity) of COA, and add HBSS equilibration solution (or other buffer suitable for cell digestion. If necessary, add EDTA to a final concentration of 0- 1mM, it is best not to exceed 2mM). The recommended trypsin concentration is about 0. 1-0.3mg/mL (concentration adjusted according to different cells), and gently mixed; this step can be operated at room temperature.
2. Use a 0.22μm filter to filter the trypsin solution and transfer it to a sterile container; This step can be operated at room temperature.
3. The filtered trypsin solution should be used directly as required on the day (for example, adding 1mL to T25 bottles, digesting cells at 37°C), and the liquid can be stored at 2-8°C for 1-2 weeks.
4. Long-term storage: If the prepared recombinant cell digestion solution needs to be stored for a long time, it can be stored stably at -25～- 15°C for 12 months.
Product usage
For cell culture
• Tissue block digestion, primary cell acquisition.
• Passage digestion of adherent cells.
• Cell culture by microcarrier method.
• Gently digesting stem cells.
• Immune cell therapy, etc.
For recombinant protein:
. Recombinant insulin production.
. Protein sequencing, peptide mapping.
. Specific proteolytic processes such asproteomics research.
Precautions
. High concentration of inorganic salts have an obvious effect on the activity.
. Avoid contact with other protein products.
. For your safety and health, please wear lab coat and gloves for operatin
