For measuring collagen secretion into the cellular medium, cells were washed with serum free medium and incubated in serum free medium for 3 h. The medium was collected and collagen was directly measured by western blot by loading 50–100 μL of the medium onto 7.5% SDS-PAGE gels27 (link),67 (link). Western blots were performed with 1:1000 dilutions of the antibodies. The antibodies used were: anti-collagen α1(I) from Rockland (600-401-103), anti-collagen α2(I) from Santa Cruz Biotechnology (sc-8786), anti-fibronectin from BD Transduction Laboratories (610523 and 610077). When cell extracts were analyzed anti-actin from Abcam (ab8227), anti-LARP6 from Novus Biologicals (H00055323) antibodies were used.
Anti collagen α1 1
Manufactured by Rockland Immunochemicals
Anti-collagen α1(I) is a laboratory reagent used in the detection and quantification of type I collagen. It is a highly specific antibody that binds to the alpha 1 chain of type I collagen, a major structural protein found in various connective tissues. This product can be utilized in immunoassays and other analytical techniques to measure the levels of type I collagen in biological samples.
Automatically generated - may contain errors
2 protocols using anti collagen α1 1
1
Measuring Secreted Collagen by Western Blot
2
Collagen Secretion Quantification
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Western blots were done with 20 μl of cellular extracts of transfected cells. The same amount was used in gel shift experiments, so that the protein expression could be directly compared to the binding activity. For measuring collagen secretion into the cellular medium, cells were washed with serum free medium and incubated in serum free medium for 3 h. The medium was collected and collagen secreted was directly measured by western blot. The antibodies used were: anti-collagen α1(I) from Rockland (600-401-103), anti-collagen α2(I) from Santa Cruz Biotechnology (sc-8786), anti-HA from Sigma (H9658), anti-calnexin and anti-fibronectin from BD Transduction Laboratories (610523 and 610077), anti-SEC61α from Upstate (07-204), anti-actin from Abcam (ab8227), anti-LARP6 from Novus Biologicals (H00055323) and anti-SEC61β from Thermo Scientific (PA3-015).
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