Anti-PHKA1 抗体 [EPR12118] (ab176338)

False colour image of Western blot: Anti-PHKA1 antibody [EPR12118] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab176338 was shown to bind specifically to PHKA1. A band was observed at 130 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in PHKA1 knockout cell line ab267337 (knockout cell lysate ab258111).

To generate this image, wild-type and PHKA1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Intracellular flow cytometric analysis of permeabilized Hela cells labeling PHKA1 with ab176338 at 1/10 dilution (red) or a rabbit IgG (negative) (green). 

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling PHKA1 with ab176338 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.