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Synthetic peptide (Human) - which represents a portion of human Mammalian Target of Rapamycin encoded in part by exons 5 and 6. (Peptide available as ab39393.)
Our Abpromise guarantee covers the use of ab2732 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/2000. Can be blocked with mTOR peptide (ab39393).|
|IHC-P||1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
mTOR protein kinase assay:
ab2732 (1:1000 dilution) immunoprecipitates were incubated at 30°C with recombinant 4E-BP1 and 32P-gATP. Autoradiograph shows 32P incorporated into 4E-BP1.
ab2732 staining rat testis sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab2732 at 1/2000 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.
Positive components in the submitted image are indicated by coloured arrowheads: red for densely positive nuclei that appear to be confined to the stem cell layer (immediately above, some spermatogonia exhibit a nuclear speckling). Yellow indicates a moderate cytoplasmic positivity, more evident in some seminiferous tubules (ST). Green for a strong apparently cytoplasmic positivity at the lumenal surface of a proportion of ST.
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