COL2A1 Antibody / Collagen alpha-1(II) chain (FY13255)

Immunohistochemical staining of Collagen Type II/COL2A1 using anti-COL2A1 antibody. Collagen Type II/COL2A1 was detected in a paraffin-embedded section of mouse knee cartilage tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-COL2A1 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of Collagen Type II/COL2A1 using anti-COL2A1 antibody. Lane 1: rat bone tissue lysates(100oC boiled for 5 minutes), Lane 2: rat bone tissue lysates(37oC incubated for 10 minutes), Lane 3: mouse bone tissue lysates(100oC boiled for 5 minutes). After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-COL2A1 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. Western blot analysis of COL2A1 in rat and mouse bone extracts using an anti COL2A1 antibody under different sample heating conditions. A predominant doublet is detected at an approximately 150 kDa in all bone lysates, running above the predicted ~142 kDa size but consistent with the reported migration of procollagen type II alpha1 chains that retain N and C propeptides and post translational modifications. Additional weaker bands above and below the major doublet likely reflect crosslinked or partially denatured collagen II species and limited degradation fragments rather than distinct alternative isoforms.

Immunohistochemical staining of Collagen Type II/COL2A1 using anti-COL2A1 antibody. Collagen Type II/COL2A1 was detected in a paraffin-embedded section of rat knee cartilage tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-COL2A1 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.