Anti-PKR antibody [YE350] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using ab32052 the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKR with Purified ab32052 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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using ab32052 the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling PKR with Purified ab32052 at 1 : 100 dilution (2.52 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using ab32052 the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PKR with Purified ab32052 at 1 : 50 dilution (5.04 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.

• IP

Lab

Immunoprecipitation - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using ab32052, the same antibody clone in a different buffer formulation.
Purified ab32052 at 1/20 dilution (1μg) immunoprecipitating PKR in Jurkat whole cell lysate.
Lane 1 (input) : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab32052 + Jurkat whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32052 in Jurkat whole cell 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 : 68 kDa
Possible degradation bands are observed between 20-30kDa.

All lanes:

Immunoprecipitation - Anti-PKR antibody [YE350] (<a href='/en-us/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>)

Predicted band size: 62 kDa

false

• WB

Supplier Data

Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using ab32052, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PKR antibody [YE350] (<a href='/en-us/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>) at 1/5000 dilution

All lanes:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 15 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: 62 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using the same antibody clone in a different buffer formulation (ab32052).

Lanes 1-3 : Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266999 (knockout cell lysate ab256900) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 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-PKR antibody [YE350] (<a href='/en-us/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

EIF2AK2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human EIF2AK2 (PKR) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-eif2ak2-pkr-knockout-a549-cell-line-ab266999'>ab266999</a>)

Lane 3:

K-562 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: 62 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using ab32052, the same antibody clone in a different buffer formulation.

Lanes 1-5 : Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-PKR antibody [YE350] (<a href='/en-us/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-eif2ak2-pkr-knockout-hela-cell-line-ab261824'>ab261824</a>)

Lane 3:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 5:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole 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: 62 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (AB239799)

This data was developed using the same antibody clone in a different buffer formulation (ab32052).

Lanes 1-3 : Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267000 (knockout cell lysate ab256901) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 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-PKR antibody [YE350] (<a href='/en-us/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

EIF2AK2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human EIF2AK2 (PKR) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-eif2ak2-pkr-knockout-a549-cell-line-ab267000'>ab267000</a>)

Lane 3:

K-562 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: 62 kDa

Observed band size: 70 kDa

false