Synthetic peptide derived from within residues 800 - 900 of Human PIWIL1.
(Peptide available as ab13827.)
Our Abpromise guarantee covers the use of ab12337 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 99 kDa (predicted molecular weight: 99 kDa).Can be blocked with Human PIWIL1 peptide (ab13827).
ab12337 detects a 99 kDa band in both mouse and rat testis lysates (positive controls). Non-specific bands at 102 kDa were also observed in mouse and rat liver lysates (negative controls).
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 10 µg/ml.|
|IP||Use at an assay dependent concentration. PubMed: 19918066|
Image courtesy of Human Protein Atlas
ab12337 staining PIWI Human female colon, showing a distinct and strong staining pattern at glandular cells. Paraffin embedded human colon tissue was incubated with ab12337 (1/200 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab12337 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
ab12337 gave a positive signal in both mouse and rat testis lysates (positive controls). Bands were also observed in mouse and rat liver lysates (negative controls) that were higher in size to that observed in testis.
ICC/IF image of ab12337 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab12337 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.