2. Metabolic Enzyme/Protease
  3. Ser/Thr Protease
  4. Chymotrypsin

Chymotrypsin  (Synonyms: EC 3.4.21.1; Chymotrypsin A)

Cat. No.: HY-108910 Purity: ≥95.0%
COA Handling Instructions

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Chymotrypsin (Chymotrypsin A) is a serine protease produced by the pancreas. Chymotrypsin cleaves protein chains at the carboxyl side of aromatic amino acids.

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Chymotrypsin Chemical Structure

Chymotrypsin Chemical Structure

CAS No. : 9004-07-3

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Other Forms of Chymotrypsin:

Chymotrypsin Related Antibodies

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Description

Chymotrypsin (Chymotrypsin A) is a serine protease produced by the pancreas. Chymotrypsin cleaves protein chains at the carboxyl side of aromatic amino acids[1][2].
In Vitro

Recommended to reconstitute in 1mM HCl at 10 mg/mL and add 2 mM calcium chloride as stabilizer. Resuspended Chymotrypsin can be stored for up to one week at 4℃.

Preparation of Protein
1. Solubilization/Denaturation: Dissolve protein in 100mM Tris-HCl, 10mM CaCl2 (pH 8.0). Proteins that are difficult to dissolve or require denaturation for efficient digestion can be solubilized in a minimum volume in a denaturant such as 6-8M urea or 6M guanidine HCl at room temperature to 37℃ for up to one hour. For some proteins, it may be beneficial to heat the sample to 60℃ over this time period (95℃ for 15-20 minutes for extreme cases).
2. Disulphide Reduction: To the dissolved protein add DTT (or β-mercaptoethanol) to a final concentration of 5mM; heat this sample at 50-60℃ for 20 minutes.
3. Alkylation: Allow the reduced protein mixture to cool to room temperature, and add iodoacetamide to a final concentration of 15mM. Incubate in the dark for 15 minutes at room temperature.
4. Finally adjust the reaction volume with 100mM Tris-HCl, 10mM CaCl2 (pH 8.0) such that the urea or guanidine concentration is 1M or less.
Enzyme Reconstitution
Dissolve Chymotrypsin in 1mM HCl. We recommend a final concentration 0.5-1 μg/μL.
Digestion
Add Chymotrypsin to a final protease:protein ratio of 1:200 to 1:20 (w/w), and incubate sample for 2-18 hours at 25℃. The reaction may be stopped, if desired, by adding 0.5% trifluoroacetic acid.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Chymotrypsin Related Antibodies
CAS No.
Appearance

Solid

Color

White to off-white

SMILES

[Chymotrypsin]
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.
Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

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Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    90% Corn Oil

    Solubility: ≥ 2.5 mg/mL (Infinity mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: ≥95.0%

COA (229 KB)

Handling Instructions (2659 KB)
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Chymotrypsin Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Chymotrypsin9004-07-3EC 3.4.21.1 Chymotrypsin ASer/Thr ProteaseSerine proteasesSerine endopeptidasesThreonine proteasesInhibitorinhibitorinhibit

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