Anti-GSTK1 antibody [EP1938Y] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GSTK1 antibody [EP1938Y] - BSA and Azide free (AB247331)

This data was developed using the same antibody clone in a different buffer formulation (ab52759).
Immunocytochemistry analysis of A549 (human lung carcinoma epithelial cell) labeling GSTK1 with purified ab52759 at 1/100 dilution (10 µg/ml). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.13 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control : PBS instead of the primary antibody.

• IP

Lab

Immunoprecipitation - Anti-GSTK1 antibody [EP1938Y] - BSA and Azide free (AB247331)

This data was developed using ab52759, the same antibody clone in a different buffer formulation.
Purified ab52759 at 1/50 dilution (2μg) immunoprecipitating GSTK1 in Human heart lysate.
Lane 1 (input) : Human heart lysate 10μg
Lane 2 (+) : ab52759 + Human heart lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab52759 in Human heart lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 25 kDa

All lanes:

Immunoprecipitation - Anti-GSTK1 antibody [EP1938Y] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-ep1938y-ab52759'>ab52759</a>)

Predicted band size: 25 kDa

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• WB

Lab

Western blot - Anti-GSTK1 antibody [EP1938Y] - BSA and Azide free (AB247331)

This data was developed using the same antibody clone in a different buffer formulation (ab52759).

Lanes 1-4 : Merged signal (red and green). Green - ab52759 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.

ab52759 Anti-GSTK1 antibody [EP1938Y] was shown to specifically react with GSTK1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266112 (knockout cell lysate ab257461) was used. Wild-type and GSTK1 knockout samples were subjected to SDS-PAGE. ab52759 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GSTK1 antibody [EP1938Y] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-ep1938y-ab52759'>ab52759</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

GSTK1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human GSTK1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gstk1-knockout-hek-293t-cell-line-ab266112'>ab266112</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 27 kDa

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