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Our Abpromise guarantee covers the use of ab37977 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration. PubMed: 24659784|
|WB||1/1000. Predicted molecular weight: 69 kDa.|
|IHC-P||1/50 - 1/100. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemical staining of SPHK2 in Human breast carcinoma tissue sections (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) with ab37977 at a dilution of 1/25. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hours at 38°C. Antigen retrieval was heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hour at 37°C. A HRP-conjugated goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.
This image is courtesy of an anonymous AbreviewBlocking Step: 5% Milk for 1 hour at 22°C.