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Full length protein corresponding to Collagen I. Collagen Type I from human and bovine placenta
The SDS-PAGE/ Coomassie stained gel of collagen I has two bands at 138K and 129K non-reduced.
It is often extremely difficult to generate antibodies with specificities to collagens due to the uninterrupted "Glycine-X-Y" triplet repeat that is a necessary part of the triple helical structure. The development of type specific antibodies is dependent on NON-DENATURED three-dimensional epitopes - this may result in diminished reactivity of some antibodies with denatured collagen or formalin-fixed, paraffin embedded tissues. Anti-Collagen antibodies have been used for indirect trapping ELISA for quantitation of antigen in serum using a standard curve, for immunoprecipitation and for native (non-denaturing, non-dissociating) PAGE and western blotting for highly sensitive qualitative analysis.
Our Abpromise guarantee covers the use of ab292 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000 - 1/50000. Detects a band of approximately 80-120 kDa. This product is not recommended for use under denaturing conditions in WB and ELISA. We would suggest testing it under native conditions.|
|ELISA||1/5000 - 1/50000.|
|IHC-Fr||1/50 - 1/200.|
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC||Use at an assay dependent concentration.|
ab292 staining Collagen I in horse bronchial fibroblast cells by Immunocytochemistry. Cells were fixed with acetone and blocking with 3% BSA was performed for 1 hour 30 minutes at 40C. Samples were incubated with primary antibody (1/500: in 3% BSA/ PBS) for 12 hours at 4°C. An FITC-conjugated goat polyclonal to rabbit IgG was used at dilution at 1/160 as secondary antibody.
ab292 at 1/200 staining rat testes tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed, before a blocking step in 1% serum. The tissue was incuabted with the primary antibody for 30 minutes at 22°C and then with the HRP conjugated goat anti-rabbit secondary.
ab292 was used at a 1:100 dilution to detect distal tubules in normal kidney tissue.
Tissue= Human lung. 1:400 dilution of ab292.
Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative.
Immunohistochemistry of ab292. Tissue: right lobe of the liver section. A:Central Vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow).
ab292 at 1:1250
Western blot of ab292. Lane 1: Human Collagen Type 1.
50 ng per lane.
Other Band(s): Collagen Type I splice variants and isoforms.
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