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AB32052

Anti-PKR antibody [YE350]

  • BOND RX™ Validated
  • RabMAb
  • Recombinant
  • KO Validated
  • What is this?

5

(1 Review)

|

(48 Publications)

Rabbit Recombinant Monoclonal PKR antibody. Suitable for ICC/IF, IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 48 publications.

View Alternative Names

PKR, PRKR, EIF2AK2, Eukaryotic translation initiation factor 2-alpha kinase 2, Interferon-inducible RNA-dependent protein kinase, P1/eIF-2A protein kinase, Protein kinase RNA-activated, Tyrosine-protein kinase EIF2AK2, p68 kinase, eIF-2A protein kinase 2, Protein kinase R

9 Images
Flow Cytometry (Intracellular) - Anti-PKR antibody [YE350] (AB32052)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PKR antibody [YE350] (AB32052)

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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [YE350] (AB32052)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed 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.

Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [YE350] (AB32052)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - 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.

Immunoprecipitation - Anti-PKR antibody [YE350] (AB32052)
  • IP

Lab

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.

All lanes:

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

Predicted band size: 62 kDa

false

Western blot - Anti-PKR antibody [YE350] (AB32052)
  • WB

Lab

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

Western blot - Anti-PKR antibody [YE350] (AB32052)
  • WB

Supplier Data

Western blot - Anti-PKR antibody [YE350] (AB32052)

Blocking buffer : 5% NFDM/TBST

All lanes:

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

Western blot - Anti-PKR antibody [YE350] (AB32052)
  • WB

Lab

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.

All lanes:

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

Predicted band size: 62 kDa

false

Western blot - Anti-PKR antibody [YE350] (AB32052)
  • WB

Lab

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

Western blot - Anti-PKR antibody [YE350] (AB32052)
  • WB

Lab

Western blot - Anti-PKR antibody [YE350] (AB32052)

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] (ab32052) 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

  • Carrier free

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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

YE350

Isotype

IgG

Carrier free

No

Reacts with

Human

Applications

Flow Cyt (Intra), IHC-P, WB, IP, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "<p><strong>For unpurified use at 1/100 - 1/500.</strong></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/20 - 1/100", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000 - 1/10000", "WB-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/20 - 1/50", "FlowCytIntra-species-notes": "<p></p>" } } }
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Product details

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

IFN-induced dsRNA-dependent serine/threonine-protein kinase that phosphorylates the alpha subunit of eukaryotic translation initiation factor 2 (EIF2S1/eIF-2-alpha) and plays a key role in the innate immune response to viral infection (PubMed : 18835251, PubMed : 19189853, PubMed : 19507191, PubMed : 21072047, PubMed : 21123651, PubMed : 22381929, PubMed : 22948139, PubMed : 23229543). Inhibits viral replication via the integrated stress response (ISR) : EIF2S1/eIF-2-alpha phosphorylation in response to viral infection converts EIF2S1/eIF-2-alpha in a global protein synthesis inhibitor, resulting to a shutdown of cellular and viral protein synthesis, while concomitantly initiating the preferential translation of ISR-specific mRNAs, such as the transcriptional activator ATF4 (PubMed : 19189853, PubMed : 21123651, PubMed : 22948139, PubMed : 23229543). Exerts its antiviral activity on a wide range of DNA and RNA viruses including hepatitis C virus (HCV), hepatitis B virus (HBV), measles virus (MV) and herpes simplex virus 1 (HHV-1) (PubMed : 11836380, PubMed : 19189853, PubMed : 19840259, PubMed : 20171114, PubMed : 21710204, PubMed : 23115276, PubMed : 23399035). Also involved in the regulation of signal transduction, apoptosis, cell proliferation and differentiation : phosphorylates other substrates including p53/TP53, PPP2R5A, DHX9, ILF3, IRS1 and the HHV-1 viral protein US11 (PubMed : 11836380, PubMed : 19229320, PubMed : 22214662). In addition to serine/threonine-protein kinase activity, also has tyrosine-protein kinase activity and phosphorylates CDK1 at 'Tyr-4' upon DNA damage, facilitating its ubiquitination and proteasomal degradation (PubMed : 20395957). Either as an adapter protein and/or via its kinase activity, can regulate various signaling pathways (p38 MAP kinase, NF-kappa-B and insulin signaling pathways) and transcription factors (JUN, STAT1, STAT3, IRF1, ATF3) involved in the expression of genes encoding pro-inflammatory cytokines and IFNs (PubMed : 22948139, PubMed : 23084476, PubMed : 23372823). Activates the NF-kappa-B pathway via interaction with IKBKB and TRAF family of proteins and activates the p38 MAP kinase pathway via interaction with MAP2K6 (PubMed : 10848580, PubMed : 15121867, PubMed : 15229216). Can act as both a positive and negative regulator of the insulin signaling pathway (ISP) (PubMed : 20685959). Negatively regulates ISP by inducing the inhibitory phosphorylation of insulin receptor substrate 1 (IRS1) at 'Ser-312' and positively regulates ISP via phosphorylation of PPP2R5A which activates FOXO1, which in turn up-regulates the expression of insulin receptor substrate 2 (IRS2) (PubMed : 20685959). Can regulate NLRP3 inflammasome assembly and the activation of NLRP3, NLRP1, AIM2 and NLRC4 inflammasomes (PubMed : 22801494). Plays a role in the regulation of the cytoskeleton by binding to gelsolin (GSN), sequestering the protein in an inactive conformation away from actin (By similarity).
See full target information EIF2AK2
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Publications (48)

Recent publications for all applications. Explore the full list and refine your search

The EMBO journal 43:3587-3603 PubMed38951609

2024

LINE-1 RNA triggers matrix formation in bone cells via a PKR-mediated inflammatory response.

Applications

Unspecified application

Species

Unspecified reactive species

Arianna Mangiavacchi,Gabriele Morelli,Sjur Reppe,Alfonso Saera-Vila,Peng Liu,Benjamin Eggerschwiler,Huoming Zhang,Dalila Bensaddek,Elisa A Casanova,Carolina Medina Gomez,Vid Prijatelj,Francesco Della Valle,Nazerke Atinbayeva,Juan Carlos Izpisua Belmonte,Fernando Rivadeneira,Paolo Cinelli,Kaare Morten Gautvik,Valerio Orlando

Science signaling 17:eadi9844 PubMed38771918

2024

Condensation of RNase L promotes its rapid activation in response to viral infection in mammalian cells.

Applications

Unspecified application

Species

Unspecified reactive species

Renee Cusic,James M Burke

Cells 13: PubMed38334619

2024

Significant Variations in Double-Stranded RNA Levels in Cultured Skin Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Shaymaa Sadeq,Suwalak Chitcharoen,Surar Al-Hashimi,Somruthai Rattanaburi,John Casement,Andreas Werner

Molecular cell 84:584-595.e6 PubMed38244546

2024

N-methyladenosine in 5' UTR does not promote translation initiation.

Applications

Unspecified application

Species

Unspecified reactive species

Ewelina Guca,Rodrigo Alarcon,Michael Z Palo,Leonardo Santos,Santiago Alonso-Gil,Marcos Davyt,Leonardo H F de Lima,Fanny Boissier,Sarada Das,Bojan Zagrovic,Joseph D Puglisi,Yaser Hashem,Zoya Ignatova

PLoS pathogens 19:e1010812 PubMed36795772

2023

NS5A domain I antagonises PKR to facilitate the assembly of infectious hepatitis C virus particles.

Applications

Unspecified application

Species

Unspecified reactive species

Shucheng Chen,Mark Harris

iScience 26:105909 PubMed36691613

2023

PKR deficiency delays vascular aging via inhibiting GSDMD-mediated endothelial cell hyperactivation.

Applications

Unspecified application

Species

Unspecified reactive species

Zhouyangfan Peng,Xiqing Tan,Liangpeng Xie,Ze Li,Sufang Zhou,Yapei Li

British journal of cancer 127:2141-2153 PubMed36216883

2022

YTHDF2-mediated FGF14-AS2 decay promotes osteolytic metastasis of breast cancer by enhancing RUNX2 mRNA translation.

Applications

Unspecified application

Species

Unspecified reactive species

Ming Zhang,Jue Wang,Yucui Jin,Que Zheng,Mengying Xing,Yuting Tang,Yunfei Ma,Lingyun Li,Bing Yao,Hao Wu,Changyan Ma

Cell death & disease 13:361 PubMed35436983

2022

circFAM120B functions as a tumor suppressor in esophageal squamous cell carcinoma via the miR-661/PPM1L axis and the PKR/p38 MAPK/EMT pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Huan Song,Dan Tian,Jian Sun,Xuhua Mao,Weimin Kong,Dian Xu,Ye Ji,Beibei Qiu,Mengyao Zhan,Jianming Wang

Oxidative medicine and cellular longevity 2022:1125932 PubMed35035655

2022

Novel lncRNA AL033381.2 Promotes Hepatocellular Carcinoma Progression by Upregulating PRKRA Expression.

Applications

Unspecified application

Species

Unspecified reactive species

Feiran Wang,Lirong Zhu,Qiang Xue,Chong Tang,Weidong Tang,Nannan Zhang,Chen Dai,Zhong Chen

Veterinary research 52:148 PubMed34930429

2021

GADD34-mediated dephosphorylation of eIF2α facilitates pseudorabies virus replication by maintaining de novo protein synthesis.

Applications

Unspecified application

Species

Unspecified reactive species

Ting Zhu,Xueli Jiang,Hangkuo Xin,Xiaohui Zheng,Xiaonuan Xue,Ji-Long Chen,Baomin Qi
View all publications
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