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Full length native protein (purified) corresponding to Human Collagen I aa 1-1464.
Database link: P02452
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 ab34710 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||1/50 - 1/200.|
|Indirect ELISA||1/5000 - 1/50000.|
|WB||1/5000 - 1/10000. This product is not recommended for use under denaturing conditions in WB, IP, and ELISA. We would suggest testing it under native conditions. Denaturing and reducing conditions will greatly diminish reactivity and selectivity of this antibody.|
|ELISA||1/5000 - 1/50000.|
Immunohistochemical analysis of formaldehyde-fixed frozen bovine tendon sections, labelling collagen I with ab34710 at a dilution of 1/500 for 2 hours at 21°C. 1% BSA was used as a blocking agent and incubated for 10 minutes at 21°C. Secondary used was a Goat anti-Rabbit Alexa Fluor® 594 conjugate used at 1/1,000.
ab34710 staining mouse kidney sections (ab4606) by IHC-Fr. Kidney tissue was cryoprotected with 30% sucrose, sectioned using a cryostat at 10 microns and mounted onto slides. After drying overnight in fume hood, sections were fixed in 4% formalin in PBS for 10 minutes. Blocking was performed with 1% BSA for 10 minutes at 21°C. Staining with ab34710 at a 1/100 dilution in TBS/BSA with 0.02% azide was performed for 2h at 21°C. A conjugated goat anti-rabbit 594 polyclonal antibody at 1/1000 was used as the secondary antibody.
ab34710 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 40°C. 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.
Lane 1: Human Collagen Type 1. 50 ng per lane.
Other Band(s): Collagen Type I splice variants and isoforms.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"