Anti-PABP antibody (ab21060)

Viral ORF57 expression is required to suppress the formation of SG during KSHV lytic infection and under arsenite stress.

ORF57 alone is sufficient to inhibit SG formation. HeLa cells transfected with an ORF57-Flag (pVM7) expressing vector or an empty vector (control) for 24 h were induced with 0.5 mM arsenite for 30 min for SG formation. The cells were stained for ORF57 (green), SG-specific TIA-1 (red) and PABPC1 (white) (ab21060) by each corresponding antibody. The nuclei were counterstained with Hoechst stain. Scale bar = 10 μm.

ICC/IF image of ab21060 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in methanol (5 min) and incubated with the antibody (ab21060, 1 µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence. 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor^® 594 WGA was used to label plasma membranes (red).

Image courtesy of Human Protein Atlas

ab21060 staining PABP in human appendix tissue. 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

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 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by ICC/IF.

HeLa cells fixed for 10 minutes with 1.9 % formaldehyde in 2 % sucrose in PBS then permeabilized 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.

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Immunocytochemical analysis of newborn porcine tracheal epithelial cells, labeling PABP with ab21060.

Cells were formaldehyde-fixed, permeabilized 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. 

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Immunocytochemical analysis of rat OLN93 oligodendroglia cells, labeling PABP with ab21060.

Cells were paraformaldehyde-fixed, permeabilized with 0.1% Triton X-100, and blocked with 10% normal goat serum for 1 hour at 20°C. Immunostaining with anti-PABP diluted 1/200 overnight at 4°C.

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