Anti-PIGK antibody [EPR17843] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling PIGK with ab201693 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing cytoplasmic staining on A549 cell line.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows :

-ve control 1 : ab201693 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HEK-293 (Human epithelial cells from embryonic kidney) cells labeling PIGK with ab201693 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PIGK with ab201693 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing cytoplasmic staining on Jurkat cell line.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows :

-ve control 1 : ab201693 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

PIGK was immunoprecipitated from 1mg of HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab201693 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab201693 at 1/2000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.

Lane 1 : HEK293 whole cell lysate10 μg (Input).
Lane 2 : ab201693 IP in HEK293 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab201693 in HEK293 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-PIGK antibody [EPR17843] (<a href='/en-us/products/primary-antibodies/pigk-antibody-epr17843-ab201693'>ab201693</a>)

Predicted band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PIGK antibody [EPR17843] (<a href='/en-us/products/primary-antibodies/pigk-antibody-epr17843-ab201693'>ab201693</a>) at 1/2000 dilution

Lane 1:

HT1080 (Human fibrosarcoma cells) cell lysate at 20 µg

Lane 2:

HEK-293 (Human epithelial cells from embryonic kidney) cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PIGK antibody [EPR17843] (<a href='/en-us/products/primary-antibodies/pigk-antibody-epr17843-ab201693'>ab201693</a>) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat kidney lysate at 10 µg

Lane 7:

C6 (Rat glial tumor cells) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-PIGK antibody [EPR17843] - BSA and Azide free (AB251354)

This data was developed using ab201693, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PIGK antibody [EPR17843] (<a href='/en-us/products/primary-antibodies/pigk-antibody-epr17843-ab201693'>ab201693</a>) at 1/2000 dilution

Lane 1:

Human pancreas lysate at 10 µg

Lane 2:

Human fetal liver lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Exposure time: 3min