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Our Abpromise guarantee covers the use of ab19347 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. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 2 µg/ml. Detects a band of approximately 130, 150 kDa.|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 19056349|
|ELISA||Use at an assay dependent concentration.|
|IHC-Fr||Use a concentration of 2 µg/ml.|
Immunohistochemical analysis of 4% formalin fixed frozen mouse stomach tissue labeling Calcium Sensing Receptor with ab19347 at 1/100 dilution overnight at 4°C, followed by fluorophore-conjugated goat anti-mouse IgG secondary antibody for 2 hours at RT.
Fresh stomach tissue was fixed in 4% formalin for 1 hour and then incubated overnight at in 25% sucrose before embedding in tissue freezing medium. Antigen retrieval was carried out on 8µm cryo-sections by incubating in sodium citrate buffer for 45 minutes at 4°C, immersing in sodium citrate buffer for 10 minutes at 100°C before washing 3 times for 5 minutes each in 1X PBS. Sections were then blocked in blocking buffer (0.3% Triton X-100 in 1X PBS containing 10% normal goat serum) for 30 minutes at RT before staining with ab19347.
Sections were also stained with DAPI nuclear stain (blue). Positive cells (green) were found at the base of the antral glands in the mouse stomach.
ab19347 at 1/100 staining rat brain (cerebral cortex) tissue sections by IHC-Fr. The tissue was paraformaldehyde fixed and blocked with serum before incubation with the primary antibody for 24 hours at 4°C. A biotinylated horse anti-mouse IgG was used as the secondary.
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