Collagenase II (Synonyms: Collagenase; collagen hydrolase) |
Catalog No.GC19588
One-Click Copy Product Info
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Collagenase Ⅱ is a matrix metalloproteinase (MMPs) used for tissue dissociation and the preparation of liver, bone, thyroid, heart, and salivary gland cells.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 9001-12-1
Sample solution is provided at 25 µL, 10mM.
Collagenase Ⅱ is a matrix metalloproteinase (MMPs) used for tissue dissociation and the preparation of liver, bone, thyroid, heart, and salivary gland cells[1,2]. Collagenase can cleave the bond between neutral amino acids and glycine in the Pro-X-Glyc-Pro sequence, which is common in collagen [3]. The enzyme activity of collagenase requires the presence of divalent cations such as Ca2+ and Zn2+ for activation[4]. Collagenase activity is inhibited by agents such as ethylenediaminetetraacetic acid (EDTA), dithiothreitol (DTT), detergents, hexachlorocyclohexane, and heavy metal ions (Hg2+, Pb2+, Cd2+) [5]. Collagenase is relatively mild and exhibits good dissociation ability at physiological temperature and pH without the need for mechanical stirring or special equipment. The optimal pH for Collagenase II is 6.3-8.5, and its enzymatic activity is ≥125 CDU/mg solid (CDU = Collagen Digestion Units).
There are approximately five common types of collagenase [6]:
(1) Collagenase Type I (GC19589): Contains relatively uniform multiple enzymatic activities (including collagenase, caseinase, clostridiopeptidase, and trypsin activities), and is commonly used for the dissociation of epithelial, lung, adipose, and adrenal tissue cells.
(2) Collagenase Type II (GC19588): Contains higher clostridiopeptidase activity and is typically used for the dissociation of liver, bone, thyroid, heart, and salivary gland tissue cells.
(3) Collagenase Type III (GC19590): Contains lower protease activity and is commonly used for the dissociation of mammary gland cells.
(4) Collagenase Type IV (GC19591): Contains lower trypsin activity and is typically used for the preparation of islet cells or experiments that require the preservation of cell surface receptors.
(5) Collagenase Type V (GC19592): Partially purified, containing higher collagenase and caseinase activities, and is typically used for the dissociation of pancreatic islet tissue and connective tissue dissociation.
References:
[1] Sekhon B S. Matrix metalloproteinases–an overview[J]. Research and Reports in Biology, 2010: 1-20.
[2] Alipour H, Raz A, Zakeri S, et al. Therapeutic applications of collagenase (metalloproteases): A review[J]. Asian Pacific Journal of Tropical Biomedicine, 2016, 6(11): 975-981.
[3] Trabelsi O, Dumas V, Breysse E, et al. In vitro histomechanical effects of enzymatic degradation in carotid arteries during inflation tests with pulsatile loading[J]. Journal of the mechanical behavior of biomedical materials, 2020, 103: 103550.
[4] Eming S, Smola H, Hartmann B, et al. The inhibition of matrix metalloproteinase activity in chronic wounds by a polyacrylate superabsorber[J]. Biomaterials, 2008, 29(19): 2932-2940.
[5] Daboor S M, Budge S M, Ghaly A E, et al. Extraction and purification of collagenase enzymes: a critical review[J]. Am. J. Biochem. Biotechnol, 2010, 6(4): 239-263.
[6] Alipour H, Raz A, Zakeri S, et al. Therapeutic applications of collagenase (metalloproteases): A review[J]. Asian Pacific Journal of Tropical Biomedicine, 2016, 6(11): 975-981.
This protocol only provides a guideline, and should be modified according to your specific needs.
1. Solution Preparation:
(1) Working Solution: Typically prepare the working solution in HBSS balanced salt solution (containing Ca2+ and Mg2+) at a concentration of 1-2 mg/mL. For tissue and cell dispersion, a working concentration of 0.5-2.5 mg/mL is recommended. For cartilage digestion, a working concentration of 1-2 mg/mL is recommended.
Note: Unused solution should be aliquoted and stored at -20°C in the dark. Thaw on ice before use and avoid repeated freeze-thaw cycles.
2. Tissue Separation Using Collagenase II:
(1) Cut the tissue into 3-4 mm pieces using sterile scalpels or scissors.
(2) Wash the tissue pieces several times with HBSS containing Ca2+ and Mg2+.
(3) Add enough HBSS containing Ca2+ and Mg2+ to immerse the tissue pieces, and add Collagenase II to the desired working concentration.
(4) Incubate at 37°C for 4-18 hours. Using a horizontal shaker and supplementing digestion with 3mM CaCl2 can improve digestion efficiency.
(5) The dispersed cells can be collected using a mesh sieve. Undigested tissue can be further digested with fresh Collagenase II working solution at 37°C.
(6) Wash the collected cells with HBSS without collagenase.
(7) Resuspend the cells in cell culture medium and seed onto culture dishes with appropriate culture medium.
3. Procedure for Obtaining Rat Synovial Cells Using Collagenase II [1] (From Literature, for Reference Only):
(1) Sample Collection: Place sterile synovial tissue from rats on ice and rinse three times with PBS to remove blood. Cut the tissue into 1 mm3 pieces in a dish.
(2) Initial Digestion: Continuously detach the tissue with 0.15% trypsin three times, each for 2 minutes. Transfer the detached cell suspension to a centrifuge tube containing serum to stop the digestion.
(3) Collagenase Digestion: Add 0.08% Collagenase II and continue detaching at 37°C for 30 minutes.
(4) Cell Collection: Combine the digestion solution and sieve it. Place the cell suspension in a 10mL centrifuge tube and centrifuge at 1000 rpm for 5 minutes. Discard the supernatant and resuspend the cells in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% fetal bovine serum (FBS).
Note: The above experimental protocol provides guidance for tissue separation using Collagenase II. Adjustments can be made according to other literature and specific experimental requirements.
References:
[1]Mao T, He C, Wu H, et al. RETRACTED ARTICLE: Silencing lncRNA HOTAIR declines synovial inflammation and synoviocyte proliferation and promotes synoviocyte apoptosis in osteoarthritis rats by inhibiting Wnt/β-catenin signaling pathway[J]. Cell Cycle, 2019, 18(22): 3189-3205.
| Cas No. | 9001-12-1 | SDF | |
| Synonyms | Collagenase; collagen hydrolase | ||
| Formula | M.Wt | ||
| Solubility | Storage | Store at -20°C | |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Biological Activity: ≥125CDU/mg solid Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 18 reference(s) in Google Scholar.)