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Collagenase I Sale

(Synonyms: 胶原蛋白酶,Collagenase; collagen hydrolase) 目录号 : GC19589

The collagenase currently has approximately four types: Type I collagenase, Type II collagenase, Type III collagenase, and Type IV collagenase.

Collagenase I Chemical Structure

Cas No.:9001-12-1

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Sample solution is provided at 25 µL, 10mM.


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Protocol for Tissue Isolation by Collagenase I:

The concentration of collagenase I storage solution is generally 1 g/mL (1000×). Small dose, frozen at -20℃ away from light. Thaw on ice before use to avoid repeated freeze-thaw. Commonly used concentrations for tissue and cell dispersion are 0.5-2.5 mg/mL, and 1-2 mg/mL for cartilage digestion.

  1. Use a sterile scalpel or scissors to cut the tissue into 3~4 mm sized tissue blocks
  2. Tissue blocks were washed several times with HBSS containing Ca2+, Mg2+.
  3. Sufficient amount of HBSS containing Ca2+, Mg2+was added to immerse the tissue mass, and collagenase I was added.
  4. Incubate at 37 ℃ for 4-18 h for digestion, and use a shaking table to improve digestion efficiency.
  5. The dispersed cells can be collected with a mesh screen for use, and the tissues that are not completely dissociated can be further incubated with fresh collagenase I working solution at 37 ℃.
  6. The collected cells were washed with collagenase-free HBSS.
  7. The cell culture medium is used to re-suspend the above cells, and the cells were inoculated on a cell culture dish with a suitable cell medium.

This protocol only provides a guideline, and should be modified according to your specific needs.

Protocol for Obtaining mouse osteoblasts using collagenase Ⅰ [1]:

  1. Draw materials:Neonatal SD rats for 1 to 3 days were immersed in 75% ethanol for 3 to 5min, and the ethanol was washed repeatedly with sterile normal saline and distilled water. The temporomandibular joint was separated on a sterile operating table, and the muscle, periosteum and fibrous connective tissue were removed, and the cartilage layer was removed to obtain subchondral bone. After three times of cleaning with PBS, the subchondral bone was cut into fragments of 1 mm× 1 mm× 1 mm, washed again with PBS until the bone was white, and an appropriate amount of double antibody was added for 10 min. Discard the liquid and place the bone piece on a sterilized filter paper to absorb the surface moisture before use.
  2. Type I collagenase digestion:Take bone pieces and add appropriate amount of pancreatic enzyme, stir and digest at 37℃ for 15 min, centrifuge at 1000 r/min for 5 min, rinse with PBS, centrifuge again, discard digestive fluid and add 0.1% type 1 collagenase 5 mL, digest for 90 min, shake once every 15 min during this period. After complete medium was added and centrifuged, proper amount of complete medium containing 15% was added, and more osteoblasts were freed by blowing up bone blocks and digested cell clumps. After cell precipitation was dispersed, the cells were transferred into 25 mL culture bottle. Change the liquid the next day, and then change the liquid once every 3 days.

This protocol only provides a guideline, and should be modified according to your specific needs.


[1]. Ni Tao, Zhang Wenli, et,al. Comparative study on the effect of modified adhesion tissue block method and modified collagenase elimination method on osteoblast proliferation [J].Journal of Kunming Medical University,2014,35(06):23-28.


The collagenase currently has approximately four types: Type I collagenase, Type II collagenase, Type III collagenase, and Type IV collagenase, which have specific applications: Type I Collagenase: Contains a relatively uniform variety of enzyme activities (including collagenase, caseinase, clostripain, and trypsin activity). It is commonly used for the preparation of epithelial cells and tissue cells. Type II Collagenase: Contains higher clostripain activity and is typically used for the preparation of cells from tissues such as heart, bone, muscle, thymus, and cartilage. Type III Collagenase: Contains lower protease activity and is frequently used for the preparation of mammary gland cells. Type IV Collagenase: Contains low trypsin activity and is usually employed for the preparation of pancreatic islet cells or for cell preparation experiments where receptor integrity needs to be maintained.

Collagenase I is classified as a proteinase derived from Bacillus histolyticus, having the capability to cleave the bonds between neutral amino acid and glycine residues within Pro-X-Gly-Pro sequences, which are prominently found in collagen proteins.

Collagenase I is relatively mild, with an initial balance of collagenase, caseinase, clostridium protease, and trypsin activity, and is well dissociated at physiological temperature and pH. The optimal pH of collagenase I is 7 ~ 8.

It has the unique ability to digest natural collagen and denatured collagen. Unique among proteases, collagenase I possesses the ability to break down the triple-helical natural collagen fibers that are abundant in connective tissues like skin, tendons, blood vessels, and bones. The proteolytic degradation facilitated by collagenase is employed in the enzymatic digestion of human tumors, mouse kidneys, adult and fetal brains, and various additional tissues, encompassing those featuring epithelial constituents. It acts exclusively on procollagen, breaking it and then being hydrolyzed by other proteases, without hydrolyzing fibrin and globulin. Detergents, hexachlorocyclohexane and heavy metal ions can reduce enzyme activity[1-5]. Collagenase I (Intravenous injection) could transiently reduceinterstitial fluid pressure (IFP) by digestion of collagen in tumors and increase tumor accumulation and gene expression of lipoplex[6].

Activity definition: One FALGPA hydrolysis unit hydrolyzes 1.0 µmole of furylacryloyl-Leu-Gly-Pro-Ala per min at pH 7.5 at 25 °C in the presence of calcium ions. One neutral protease unit hydrolyzes casein to produce color equivalent to 1.0 µmole tyrosine per 5 hr at pH 7.5 at 37℃. One clostripain unit hydrolyzes 1.0 µmole of BAEE per min at pH 7.6 at 25℃ in the presence of DTT.



胶原酶I相对温和,胶原酶、酪蛋白酶、梭状芽孢杆菌蛋白酶和胰蛋白酶活性初始平衡,在生理温度和pH下解离良好,最佳pH为7 ~ 8。


活性定义:1个FALGPA水解单元在钙离子存在下,在pH 7.5、25℃下每分钟水解1.0 µmol furylacryloyl-Leu-Gly-Pro-Ala。一个中性蛋白酶单位在37℃、pH 7.5条件下每5小时水解酪蛋白产生相当于1.0 µmol酪氨酸的颜色。在DTT存在下,一个clostripain单位在25℃、pH 7.6条件下每分钟水解1.0 µmol BAEE。

[1]. Pilcher BK, Sudbeck BD, et,al. Collagenase-1 and collagen in epidermal repair. Arch Dermatol Res. 1998 Jul;290 Suppl:S37-46. doi: 10.1007/pl00007452. PMID: 9710382.
[2]. Ohbayashi N, Yamagata N, et,al.Enhancement of the structural stability of full-length clostridial collagenase by calcium ions. Appl Environ Microbiol. 2012 Aug;78(16):5839-44. doi: 10.1128/AEM.00808-12. Epub 2012 Jun 8. PMID: 22685155; PMCID: PMC3406112.
[3]. Brandhorst H, Brandhorst D, et,al. Successful human islet isolation utilizing recombinant collagenase. Diabetes. 2003 May;52(5):1143-6. doi: 10.2337/diabetes.52.5.1143. PMID: 12716744.
[4]. Hamzeh Alipour, et al. Therapeutic applications of collagenase (metalloproteases): A review. Asian Pac J Trop Biomed, 2016, 6(11): 975-981.
[5]. O'Flanagan CH, Campbell KR, et,al. Dissociation of solid tumor tissues with cold active protease for single-cell RNA-seq minimizes conserved collagenase-associated stress responses. Genome Biol. 2019 Oct 17;20(1):210. doi: 10.1186/s13059-019-1830-0. PMID: 31623682; PMCID: PMC6796327.
[6]. Kato M, Hattori Y, Kubo M, Maitani Y. Collagenase-1 injection improved tumor distribution and gene expression of cationic lipoplex. Int J Pharm. 2012 Feb 28;423(2):428-34. doi: 10.1016/j.ijpharm.2011.12.015. Epub 2011 Dec 17. PMID: 22197775.

Chemical Properties

Cas No. 9001-12-1 SDF
别名 胶原蛋白酶,Collagenase; collagen hydrolase
分子式 分子量
溶解度 >5mg/mL in water 储存条件 Store at -20°C
General tips 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。
储备液的保存方式和期限:-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。
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动物体内配方计算器 (澄清溶液)

给药剂量 mg/kg 动物平均体重 g 每只动物给药体积 ul 动物数量
% DMSO % % Tween 80 % saline