Anti-PHKA1 antibody [EPR12118] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHKA1 antibody [EPR12118] - BSA and Azide free (AB249913)

This data was developed using ab176338, the same antibody clone in a different buffer formulation.

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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PHKA1 antibody [EPR12118] - BSA and Azide free (AB249913)

This data was developed using ab176338, the same antibody clone in a different buffer formulation.

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

• WB

Lab

Western blot - Anti-PHKA1 antibody [EPR12118] - BSA and Azide free (AB249913)

This data was developed using ab176338, the same antibody clone in a different buffer formulation. 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.

Lanes 1 - 2:

Western blot - Anti-PHKA1 antibody [EPR12118] - BSA and Azide free (ab249913) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-PHKA1 antibody [EPR12118] (<a href='/en-us/products/primary-antibodies/phka1-antibody-epr12118-ab176338'>ab176338</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PHKA1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PHKA1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-phka1-knockout-hek-293t-cell-line-ab267337'>ab267337</a>)

Lane 2:

Western blot - Human PHKA1 knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-phka1-knockout-hek-293t-cell-lysate-ab258111'>ab258111</a>)

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 130 kDa

false

• WB

Supplier Data

Western blot - Anti-PHKA1 antibody [EPR12118] - BSA and Azide free (AB249913)

This data was developed using ab176338, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PHKA1 antibody [EPR12118] (<a href='/en-us/products/primary-antibodies/phka1-antibody-epr12118-ab176338'>ab176338</a>) at 1/1000 dilution

Lane 1:

A431 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

Human skeletal muscle lysate at 10 µg

Predicted band size: 137 kDa

false