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Full length native protein (purified) corresponding to Human Collagen VI aa 1-1028. Collagen Type VI from human and bovine placenta.
Database link: P12109
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.
Our Abpromise guarantee covers the use of ab6588 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/250 - 1/500.|
|ELISA||1/4000 - 1/8000.|
|IHC-Fr||1/50 - 1/200.|
|IP||Use at an assay dependent concentration.|
|WB||1/5000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 108 kDa). This product is not recommended for use under denaturing conditions in WB, IP, and ELISA. We would suggest testing it under native conditions.|
|IHC-P||Use at an assay dependent concentration. PubMed: 18065394|
Immunohistochemical analysis of 10% buffered formalin-fixed swine lung tissue sections, labelling Collagen VI with ab6588 at a dilution of 1/400 incubated for 12 hours at 4°C. Antigen retrival was with 10mM sodium citrate buffer pH 6.0 (heat mediated). Blocking was with 5% serum incubated for 1 hour at 21°C. The secondary was a Donkey anti-Rabbit polyclonal Alexa Fluor® 647 conjugtate undiluted.
ab6588 staining (green) Collagen VI in a primary cell culture from Mouse retinal pigment epithelium by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody 1/500 (1% goat serum, 0.1% TX100, 1XPBS) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat polyclonal to rabbit IgG (ab150077), dilution 1/500, was used as secondary antibody. Nuclei were counterstained with DAPI (blue).
ab6588 staining Collagen VI in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed in paraformaldehyde and an antigen retrieval step was performed using citrate buffer. Samples were then blocked with 10% serum for 30 minutes at 20°C and then incubated with ab6588 at a 1/50 dilution for 1 hour at 37°C. The secondary used was an Alexa-Fluor 555 conjugated goat anti-rabbit polyclonal (ab150078), used at a 1/500 dilution.
ab6588 at 1/400 dilution staining Collagen VI in Human placenta (Top) with red staining of stromal and extracellular spaces, and in testis (Right) with staining of extracellular spaces between seminiferous tubules. Slides were steamed in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes for antigen retrieval.
Immunohistochemical analysis of Formaldehyde fixed human colon tissue sections labelling Collagen VI with ab6588 at a dilution of 1/200. The secondary antibody used was HRP conjugated rabbit polyclonal secondary. Antigen retrieval was heat mediated.
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