Trypsin is a major proteolytic enzyme, synthesized as a preproenzyme by pancreas and is stored as proenzyme trypsinogen in secretory granules. Trypsin belongs to the family of serine proteases that are characterized by the catalytic triad His57, Asp102 and Ser195. Trypsin is routinely used in proteomics research for peptide mapping and protein sequence work due to its highly specific cleavage resulting in a limited number of tryptic peptides. Trypsin is a pancreatic serine endoprotease which hydrolyzes peptide bonds specifically at the carboxyl side of arginine and lysine residues. The rate of hydrolysis is slower if an acidic residue is on either side of the cleavage site and cleavage may not occur if a proline residue is on the carboxyl side. The enzyme also exhibits esterase and amidase activities.
Usage
For cell culture
1. Tissue block digestion, primary cell acquisition;
2. Passage digestion of adherent cells;
3. Cell culture by microcarrier method;
4. Gently digesting stem cells;
5. Immune cell therapy, etc.
For recombinant protein
1. Recombinant insulin production;
2. Protein sequencing, peptide mapping;
3. Specific proteolytic processes such as proteomics research.
Features
HPLC/SDS-PAGE > 97%
Microbial testing
WFI/PW testing
Mycoplasma testing
Extremely low endotoxin level
Host cell DNA and protease residue testing
Heavy metal residue testing
Tag-free
Comprehensive regulatory support file
Batch to batch consistency
One USP Unit will produce an A253nm of 0.003 per minute with BAEE as substrate at pH7.6 at 25℃ in a reaction volume of 3.2mL
1. Solution preparation: Melt Recombinant Trypsin at room temperature, take an appropriate amount of trypsin according to the protein content 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 2 mM). 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 0.22 μm 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 phosphatase cell digestion solution needs to be stored for a long time, it can be stored stably at -20 °C for 12 months.
20MM CaCl2，10MM HCl，pH 2±0.5
For cell culture
1. Tissue block digestion, primary cell acquisition;
2. Passage digestion of adherent cells;
3. Cell culture by microcarrier method;
4. Gently digesting stem cells;
5. Immune cell therapy, etc.
For recombinant protein
1. Recombinant insulin production;
2. Protein sequencing, peptide mapping;
3. Specific proteolytic processes such as proteomics research.
Quality Standards
| Items | Standards |
Appearence | Sterile Filtered White lyophilized (freeze-dried) powder and sterile clear, colorless liquid |
Molecule Weight | 23KDa |
Purity | β-trypsin≥70%，α-trypsin≤20% |
Bioactivity | ≥ 3800U/mg (One USP Unit will produce an A253nm of 0.003 per minute with BAEE as substrate at pH7.6 at 25℃ in a reaction volume of 3.2mL) |
Endotoxin | ＜0.5 EU/100U as determined by TAL method |
Sterility | Negative |
Mycoplasma | Negative |
Host Cell DNA | <0.02 ng/μg of protein tested by DNA Fluorescent Staining method. |
Host Cell Protein | <0.5 ng/μg of protein tested by ELISA |
外观：无色澄清透明 |
目测 |
鉴别：阳性 |
中国药典2020版 通则（3401） |
宿主蛋白残留 ≤ 0.005% |
中国药典2020版 通则（0731） |
pH值 ：7.0 - 9.0 |
中国药典2020版 通则（0631） |
酶活 ≥ 250kU/mL |
在37℃，pH8的反应条件，2.625mL的反应体系中，在30分钟内使ΔA260吸收值变化 1.0（完全消化37μg鲑鱼精DNA成为寡核苷酸）所用的酶量定义为1个酶活单位（U） |
纯度 ≥ 99% |
中国药典2020版 通则（0541）、 SEC-HPLC法 |
蛋白酶活性：无活性 |
1 kU/ml 酶，与底物37℃ 孵育60 min |
内毒素 ≤ 0.01 EU/kU |
中国药典2020版 通则（1143） |
微生物：不得检出 |
中国药典2020版 通则（1101） |
病原体：阴性 |
中国药典2020版 通则（3306） |
| 支原体：阴性 |
支原体检测试剂盒，PCR法 |
重金属残留 ≤ 10 ppm |
中国药典2020版 通则（0821） |
Storage Class Code | WGK | Flash Point（F) |
8A - Combustible, corrosive hazardous materials | WGK 3 | Not Applicable |
Flash Point（C) | Personal Protection | |
Not Applicable | Eye shields, gloves | |
COMPOSITION E.Coli Recombinant Trypsin（Porcine Sequence) | |
MOLECULE WEIGHT APPROX. 23kDa | |
Specs 1mL/1mg-1000mg | APPEARENCE Sterile Filtered White lyophilized (freeze-dried) powder and sterile clear, colorless liquid |
ENDOTOXINES < 0.5 EU/100U | Host Cell DNA Residue <0.02 ng/μg |
PURITY β-trypsin≥70%，α-trypsin≤20% | Host Cell Protein <0.5 ng/μg |
| BIOACTIVITY ≥ 3800U/mg | |
FORMULATION 20MM CaCl2，10MM HCl，pH2±0.5 | |
USAGE For Research Use or Further Manufacture Use | |
RECONSITITUION Before use this product, please read the direction below carefully. 1. This vial must be briefly centrifuged prior to opening to bring the contents to the bottom. 2. Reconstitute in a sterile aqueous buffer to an appropriate concentration. 3. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions. | |
STABILITY & STORAGE 12 months from date of receipt, the product should be stored ≤-20°C. Please avoid repeated freeze-thaw cycles after reconstitution. | |
| SHIPPING The product is shipped with dry ice(liquid) or wet ice (powder). Upon receipt, store it immediately at the temperature recommended. | |
Specs
FAQs
Composition | Recombinant Trypsin (porcine sequence) |
Cat No. | GB04.1/GB04.2/GB04.3/GB04.4/GB04.5 |
| Appearence | Sterile filtered clear liquid/Sterile filtered white lyophilized (freeze-dried) powder. |
| Host | E.Coli |
| Molecule Weight | 23.29KDa |
| Biological Activity | ≥ 3800U/mg (One USP Unit will produce an A253nm of 0.003 per minute with BAEE as substrate at pH7.6 at 25℃ in a reaction volume of 3.2mL) |
| Purity | β-trypsin≥70%，α-trypsin≤20% |
| Formulation | 20MM CaCl2，10MM HCl，pH 2±0.5 |
| Usage | This GMP product can be for research use or further manufacturing use |
| Endotoxin | < 0.05 EU/100U by LAL |
| Mycoplasma | Negative |
In Vitro Virus Assay | Negative |
| Host Cell DNA | Negative |
Host Cell Protein | Negative |
| Sterility | Negative |
Reconstitution | Before use this product, please read the direction below carefully. 1. This vial must be briefly centrifuged prior to opening to bring the contents to the bottom. 2. Reconstitute in a sterile aqueous buffer to an appropriate concentration. 3. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions. |
| Stability&Storage | 12 months from date of receipt, the product should be stored ≤-20°C. Please avoid repeated freeze-thaw cycles after reconstitution. |
Shipping | The product is shipped with dry ice(liquid) or wet ice (powder). Upon receipt, store it immediately at the temperature recommended. |
| Usage |
For cell culture 1. Tissue block digestion, primary cell acquisition; 2. Passage digestion of adherent cells; 3. Cell culture by microcarrier method; 4. Gently digesting stem cells; 5. Immune cell therapy, etc. For recombinant protein 1. Recombinant insulin production; 2. Protein sequencing, peptide mapping; 3. Specific proteolytic processes such as proteomics research. |
| Solution Preparation |
1. Melt BETAGENE™ rPT at room temperature, take an appropriate amount of rPT according to the protein content 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. Do not exceed 2 mM). The recommended rPT concentration is about 0.1-0.3mg/mL (concentration adjusted according to different cells); 2. Filter rPT solution with 0.22 μm and transfer it to a sterile container; 3. The filtered rPT 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. Store the digestion solution prepared at -20 °C for long term. |
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