Anti-GSTK1 antibody [EPR1939] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GSTK1 antibody [EPR1939] - BSA and Azide free (AB248711)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GSTK1 with Purified ab134173 at 1/30 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134173).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GSTK1 antibody [EPR1939] - BSA and Azide free (AB248711)

Immunocytochemistry/ Immunofluorescence analysis of 293T (Human embryonic kidney epithelial cell) cells labeling GSTK1 with Purified ab134173 at 1/50 dilution (5.92 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134173).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSTK1 antibody [EPR1939] - BSA and Azide free (AB248711)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling GSTK1 with Purified ab134173 at 1/300 dilution (0.99 μg/mL). Heat mediated antigen retrieval using Bond™™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134173).

• IP

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Immunoprecipitation - Anti-GSTK1 antibody [EPR1939] - BSA and Azide free (AB248711)

Purified ab134173 at 1/30 dilution (2ug) immunoprecipitating GSTK1 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate (10μg)
Lane 2 (+) : ab134173 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab134173 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 25 kDa
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134173).

All lanes:

Immunoprecipitation - Anti-GSTK1 antibody [EPR1939] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-epr1939-ab134173'>ab134173</a>)

Predicted band size: 25 kDa

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• WB

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Western blot - Anti-GSTK1 antibody [EPR1939] - BSA and Azide free (AB248711)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134173).

All lanes:

Western blot - Anti-GSTK1 antibody [EPR1939] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-epr1939-ab134173'>ab134173</a>) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

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• WB

Lab

Western blot - Anti-GSTK1 antibody [EPR1939] - BSA and Azide free (AB248711)

This data was developed using the same antibody clone in a different buffer formulation (ab134173).

Lanes 1-4 : Merged signal (red and green). Green - ab134173 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.

ab134173 Anti-GSTK1 antibody [EPR1939] 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. ab134173 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 [EPR1939] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-epr1939-ab134173'>ab134173</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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