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Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Human PABP.
(Peptide available as ab23710.)
Our Abpromise guarantee covers the use of ab21060 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.|
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa.|
|IHC-P||1/40. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 1 µg/ml.|
Immunocytochemical analysis of rat OLN93 oligodendroglia cells, labeling PABP with ab21060. Cells were paraformaldehyde fized, permeabilized with 0.1% Triton X100, and blocked with 10% Normal Goat Serum for 1 hour at 20°C. Immonstaining with anti-PABP diluted 1/200 overnight at 4°C.
ab21060 detects a band of ~ 75 kDa in HeLa whole cell lysate. A 75 kDa band is also seen in Jurkat and A431 whole cell lysates (data not shown). PABP is reported to have run at an apparent molecular mass of 73 kDa on SDS-PAGE. We believe the band we see at 75 kDa represents PABP.
ab21060 at 1/50 staining human HeLa cells by ICC/IF. HeLa cells fixed for 10 minutes with 1.9 % formaldehyde in 2 % sucrose in PBS then permeabilised for 10 minutes with 0.5% NP40 in PBS-10% sucrose. The cells were then blocked in horse serum and incubated with the antibody for 1 hour. An Alexa Fluor ® 555 conjugated donkey anti-rabbit antibody was used as the secondary.
Immunocytochemical analysis of newborn porcine tracheal epithelial cells, labeling PABP with ab21060. Cells were formaldehyde fixed, premeabilized in cold methanol, and blocked with 5% horse serum in PBS for 40 minutes at 23°C. Immunostaining with primary anti-PABP diluted 1/250 for 12 hours at 4°C.
Image courtesy of Human Protein Atlas
ab21060 staining PABP in Human appendix. The paraffin embedded tissue was incubated with ab21060 (1/40 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab21060 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.or