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Our Abpromise guarantee covers the use of ab5097 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. PubMed: 17028989|
|ICC/IF||Use a concentration of 5 µg/ml.|
ICC/IF image of ab5097 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab5097 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab5097 staining CTGF in human heart tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in buffer and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/50 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.
ab5097 staining CTGF in Human Hela, cervix cancer cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 1% normal goat serum for 30 minutes. Samples were incubated with primary antibody (1/750) for 4 hours at 20°C. A Cy3®-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody (1/800).
ab5097 staining CTGF in Pig skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 2 hours at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in blocking buffer) for 24 hours at 4°C. A BHRP-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.
Western blot, on recombinant Human CTGF produced in E. coli, using ab5097 at 1/500.
Lane 1: 0.01µg Human CTGF
Lane 2: 0.01µg Human CTGF + CTGF blocking peptide (ab13741)
Lane 3: 0.1µg Human CTGF
Lane 4: 0.1µg Human CTGF + CTGF blocking peptide (ab13741)
Secondary ab: Rabbit polyclonal to Goat IgG HRP ab6741 (1/5000)
Exposure time: 10 seconds
Expected molecular weight: 38.3 kDa
CTGF Blocking peptide (ab13741) used at 1µg/ml.
Expected molecular weight: 38kDa
Not yet tested against endogenous CTGF
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