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Our Abpromise guarantee covers the use of ab59509 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||Use at an assay dependent concentration. Use under non reducing condition. Predicted molecular weight: 37 kDa.|
|IP||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
ab59509 staining CD82 in Rat placenta tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilsed with 0.3% Triton X-100 and blocked with 3% serum for 60 minutes at 23°C; antigen retrieval was by heat mediation in 10mM citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 12 hours at 4°C. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/100) was used as the secondary antibody.
ab59509 staining CD82 (left) in Human peripheral blood neutrophils by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 1% BSA for 30 minutes at 22°C. Samples were incubated with primary antibody (1/100 in PBS + 0.1% Triton-X 100 + 1% BSA) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody. Middle - LFA 1a, Right - merge.
Overlay histogram showing HeLa cells stained with ab59509 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab59509, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG, H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.