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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.|
|IHC||Use at an assay dependent concentration. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.|
|ELISA||1/5000 - 1/50000.|
|IHC-P||Use at an assay dependent concentration.|
ab34710 staining Collagen I in Pig small intestines tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
ab34710 at 1/500 staining human adrenal tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was blocked and then incubated with the primary antibody for 30 minutes at 22°C and then with an HRP conjugated goat anti-rabbit secondary antibody.
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 was used at a 1:100 dilution to detect distal tubules in normal kidney tissue.
ab34710 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.
Tissue= Human lung. 1:400 dilution of ab34710. Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative.
Immunohistochemistry of ab34710. 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).
ab34710 at 1:1250
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"