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Full length native protein (purified) corresponding to Human Collagen IV aa 1-1669. Collagen Type IV from human and bovine placenta. The immunogen maintains the native conformation of the protein.
Database link: P02462
At least 11 genetically distinct gene products are collectively referred to as 'collagen types' or other proteins and proteoglycans of the extracellular matrix. In humans, collagens are composed of about 20 unique protein chains which under go various types of post-translational modifications and are ultimately assembled into a triple helix. This results in great diversity between collagen types. Collagens are highly conserved throughout evolution and are characterized by an uninterrupted "Glycine-X-Y" triplet repeat that is a necessary part of the triple helical structure. For these reasons it is often extremely difficult to generate antibodies with specificities to collagens. The development of type specific antibodies is dependent on NON-DENATURED three-dimensional epitopes. This preparation results in a native conformation of the protein.
This antibody is well suited to detect extracellular matrix proteins in normal as well as disease state tissues. Disruption of tissue organization is the hallmark of neoplasia. Malignant lesions can be distinguished from benign by examining the breakdown of basement membranes and loss of 3-dimensional architecture. Malignant cells are presumed to use matrix metalloproteases to degrade barriers created by the extracellular matrix which then allows metastasis to occur. Collagenases, stomelysins and gelatinases can collectively degrade all of the various components of the extracellular matrix, including fibrillar and non-fibrillar collagens and basement membrane glycoproteins.
Our Abpromise guarantee covers the use of ab6586 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||1/5000 - 1/50000.|
|IHC-Fr||1/50 - 1/200.|
|WB||1/1000 - 1/10000. Use under non reducing condition. This product is not recommended for use under denaturing conditions in WB, IP, and ELISA. We would suggest testing it under native conditions.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 19933193|
|IHC-P||1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-FrFl||Use at an assay dependent concentration.|
|IHC-FoFr||Use at an assay dependent concentration.|
ab6586 staining Collagen IV in Dog liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a 10mM citrate buffer, pH 6.0. Samples were incubated with primary antibody (1/200 in PBS + 1x casein) for 1 hour at 37°C. An undiluted HRP-conjugated Horse anti-rabbit IgG polyclonal was used as the secondary antibody.
Immunocytochemical analysis of Mouse 3T3 cell labeling Collagen IV with ab6586 at 1/250 dilution
This image is courtesy of an anonymous AbreviewBlocked with 5% milk
ab6586 staining rat brain tissue sections (ab4616) by IHC-Fr. Sections were acetone fixed and blocked with 1% serum for 20 minutes at 25°C. The primary antibody was diluted 1/300 and incubated with the sample for 30 minutes at 25°C. A biotinylated goat anti-rabbit IgG antibody was used as the secondary.
ab6586 staining Collagen IV in Human corneal epithelial tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was enzymatic using trypsin. Samples were incubated with primary antibody (1/500 inPBS + 2%NGS) for 18 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal (1/500) (ab150077) was used as the secondary antibody.
This image is courtesy of an anonymous AbreviewBlocked with 5% BSA for 1 hour at 25°C.
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