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Our Abpromise guarantee covers the use of ab2365 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 85 kDa.|
|IHC-P||Use at an assay dependent concentration. This antibody may be diluted to a titer of 1:100-1:200 in an ABC method. We suggest an incubation period of 30 minutes at room temperature. However, depending upon the fixation conditions and the staining system employed, optimal incubation conditions and antibody dilutions should be determined by the user. Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
|ICC/IF||1/100. (see Abreview).|
|IP||Use a concentration of 5 µg/ml.|
ab2365 at 1/100 staining rat gastric epithelial cells by ICC/IF. The cells were fomaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 2 hours. A FITC conjugated goat anti-rabbit IgG was used as the secondary. Nuclei were counterstained with propidium iodide.
The image shows the staining of gastric epithelial cells (left hand panel) and the staining of embyonic fibroblasts (negative control, right hand panel).
Beta Catenin was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to beta Catenin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, A431 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2365.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 85kDa; beta Catenin.
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