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Collagen is a protein that strengthens and supports many tissues in the body. Type I collagen (Collagen I) is most abundant, encoded by the COL1A1 gene. It has important roles in tissue structure, rigidity, and elasticity.
Collagen I can be a challenging target to work with in western blot.
Collagen I is the major protein component of the extracellular matrix in bone, skin and tendon, mainly secreted by osteoblasts, dermal fibroblasts and tenocytes. Proper collagen isolation method is critical for western blot success.
|Electrophoresis||Use a 6% acrylamide gel for collagen I.|
|Transferring||It is preferred to add SDS to a final concentration of 0.1% in the transfer buffer for large proteins.|
|To determine if the transfer is successful by visualization of proteins in membranes using Ponceau S.|
|Maximizing signal||Acid or enzyme treatment with pepsin is a better method to isolate collagen.|
|Continuous refrigeration throughout collagen extraction is important to avoid degradation and denaturation.|
Take care with pH, temperature, and concentration to avoid collagen polymerization:
Human stomach, skin and adrenal gland tissue lysates
Find full information on working with Collagen I: