Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [YE350] to PKR
Suitable for: Flow Cyt, WB, IHC-P, IP, ICC
Knockout validated
Reacts with: Human

Product name

Anti-PKR antibody [YE350]
See all PKR primary antibodies
Description

Rabbit monoclonal [YE350] to PKR
Host species

Rabbit
Specificity

ab32052 recognises Protein kinase R (PKR). It does not cross-react with other GCN2 family members.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human PKR aa 50-150. The exact sequence is proprietary.
Database link: P19525
Positive control
- WB: Jurkat, A549, K562, HAP1, HepG2, and HeLa cell lysates. IP: Jurkat cell lysat; IHC: Human cerebrum tissue; ICC: MCF7 cells; Flow Cyt: HeLa cells.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

YE350
Isotype

IgG
Research areas

Western blot - Anti-PKR antibody [YE350] (ab32052)

All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 5 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa
why is the actual band size different from the predicted?

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.
Immunocytochemistry - Anti-PKR antibody [YE350] (ab32052)

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.
Immunohistochemistry paraffin embedded sections - Anti-PKR antibody [YE350] (ab32052)

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.
Western blot - Anti-PKR antibody [YE350] (ab32052)

Anti-PKR antibody [YE350] (ab32052) at 1/5000 dilution (Purified) + Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Flow Cytometry - Anti-PKR antibody [YE350] (ab32052)

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).
Immunoprecipitation - Anti-PKR antibody [YE350] (ab32052)

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.
Western blot - Anti-PKR antibody [YE350] (ab32052)

All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : EIF2AK2 knockout A549 cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

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.
Western blot - Anti-PKR antibody [YE350] (ab32052)

All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : EIF2AK2 knockout A549 cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

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.
Western blot - Anti-PKR antibody [YE350] (ab32052)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: EIF2AK2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: K652 whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab32052 was shown to specifically react with EIF2AK2 when EIF2AK2 knockout samples were used. Wild-type and EIF2AK2 knockout samples were subjected to SDS-PAGE. Ab32052 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

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