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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human TRA2B.
(Peptide available as ab32689.)
Our Abpromise guarantee covers the use of ab31353 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|CLIP||Use at an assay dependent concentration. PubMed: 22194695|
|IHC-P||1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 33 kDa). Although the predicted molecular mass of TRA2B is ~33 kDa, the available literature describes TRA2B running at ~40 kDa in SDS-PAGE|
|ICC/IF||Use a concentration of 1 µg/ml.|
ICC/IF image of ab31353 stained HeLa cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab31353 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Image courtesy of Human Protein Atlas . ab31353 staining TRA2B in human pancreas, showing a distinct and strong nuclear staining pattern in both exocrine pancreas and islet cells. Paraffin embedded human pancreas tissue was incubated with ab31353 (1/300 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab31353 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues. Further results for this antibody can be found at www.proteinatlas.org