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AB220801

Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free

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(26 Publications)

Rabbit Recombinant Monoclonal PKR phospho T446 antibody. Carrier free. Suitable for IP, WB and reacts with Human samples. Cited in 26 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

2 Images
Immunoprecipitation - Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free (AB220801)
  • IP

Supplier Data

Immunoprecipitation - Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free (AB220801)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32036).

PKR was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A(100nM 30min) whole cell lysate 10 μg with ab32036 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A(100nM 30min) whole cell lysate 10 μg

Lane 2 : ab32036 IP in HeLa treated with Calyculin A(100nM 30min) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab32036 in HeLa treated with Calyculin A(100nM 30min) whole cell lysate

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

Lower bands could be cleavage form. (PMID : 28702377)

All lanes:

Immunoprecipitation - Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free (ab220801)

Predicted band size: 62 kDa

false

Western blot - Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free (AB220801)
  • WB

Lab

Western blot - Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free (AB220801)

Blocking/Diluting buffer : 5% NFDM/TBST

Positive control : Rabbit monoclonal [Y117] to PKR (ab32506).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32036).

All lanes:

Western blot - Anti-PKR (phospho T446) antibody [E120] (<a href='/en-us/products/primary-antibodies/pkr-phospho-t446-antibody-e120-ab32036'>ab32036</a>) at 1/2000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma) whole cell lysate at 15 µg

Lane 2:

HeLa (human cervix adenocarcinoma) treated with 100 ng/mL Calyculin A whole cell lysate at 15 µg

Lane 3:

HeLa (human cervix adenocarcinoma) treated with 100 ng/mL Calyculin A followed by 20 ng/mL TNF alpha 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: 68 kDa

false

Exposure time: 10s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

E120

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, IP

applications

Immunogen

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

Reactivity data

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Product details

ab220801 is the carrier-free version of ab32036.

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.

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

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Product protocols

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

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 phospho T446

Publications (26)

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

PLoS pathogens 12:e1005982 PubMed27783669

2016

Middle East Respiratory Coronavirus Accessory Protein 4a Inhibits PKR-Mediated Antiviral Stress Responses.

Applications

WB

Species

Human

Huib H Rabouw,Martijn A Langereis,Robert C M Knaap,Tim J Dalebout,Javier Canton,Isabel Sola,Luis Enjuanes,Peter J Bredenbeek,Marjolein Kikkert,Raoul J de Groot,Frank J M van Kuppeveld

Journal of virology 90:6140-7 PubMed27122577

2016

NSs Virulence Factor of Rift Valley Fever Virus Engages the F-Box Proteins FBXW11 and β-TRCP1 To Degrade the Antiviral Protein Kinase PKR.

Applications

Unspecified application

Species

Unspecified reactive species

Markus Kainulainen,Simone Lau,Charles E Samuel,Veit Hornung,Friedemann Weber

Retrovirology 11:93 PubMed25389016

2014

ADAR1 enhances HTLV-1 and HTLV-2 replication through inhibition of PKR activity.

Applications

Unspecified application

Species

Unspecified reactive species

Anne Cachat,Sandrine Alais,Sébastien Alain Chevalier,Chloé Journo,Floriane Fusil,Hélène Dutartre,Adrien Boniface,Nga Ling Ko,Antoine Gessain,François-Loïc Cosset,Rodolphe Suspène,Jean-Pierre Vartanian,Renaud Mahieux

Oncotarget 5:7563-74 PubMed25149538

2014

βTrCP controls the lysosome-mediated degradation of CDK1, whose accumulation correlates with tumor malignancy.

Applications

Unspecified application

Species

Unspecified reactive species

Joaquín Herrero-Ruiz,Mar Mora-Santos,Servando Giráldez,Carmen Sáez,Miguel A Japón,Maria Tortolero,Francisco Romero

Journal of cell science 127:2471-82 PubMed24659800

2014

IPS-1 plays an essential role in dsRNA-induced stress granule formation by interacting with PKR and promoting its activation.

Applications

WB

Species

Unspecified reactive species

Peifen Zhang,Yuye Li,Jun Xia,Junfang He,Jieying Pu,Jiong Xie,Siyu Wu,Lianqiang Feng,Xi Huang,Ping Zhang

PLoS pathogens 10:e1004002 PubMed24626510

2014

Adaptive gene amplification as an intermediate step in the expansion of virus host range.

Applications

WB

Species

Human

Greg Brennan,Jacob O Kitzman,Stefan Rothenburg,Jay Shendure,Adam P Geballe

Journal of immunology (Baltimore, Md. : 1950) 192:3753-64 PubMed24623135

2014

NF90 exerts antiviral activity through regulation of PKR phosphorylation and stress granules in infected cells.

Applications

WB

Species

Unspecified reactive species

Xi Wen,Xiaofeng Huang,Bobo Wing-Yee Mok,Yixin Chen,Min Zheng,Siu-Ying Lau,Pui Wang,Wenjun Song,Dong-Yan Jin,Kwok-Yung Yuen,Honglin Chen

PloS one 9:e87517 PubMed24475301

2014

Reporters transiently transfected into mammalian cells are highly sensitive to translational repression induced by dsRNA expression.

Applications

WB

Species

Human

Jana Nejepinska,Radek Malik,Susan Wagner,Petr Svoboda

Journal of virology 88:456-68 PubMed24155404

2013

Measles virus C protein impairs production of defective copyback double-stranded viral RNA and activation of protein kinase R.

Applications

WB

Species

Unspecified reactive species

Christian K Pfaller,Monte J Radeke,Roberto Cattaneo,Charles E Samuel

Journal of virology 87:13386-96 PubMed24089560

2013

Alpha interferon restricts human T-lymphotropic virus type 1 and 2 de novo infection through PKR activation.

Applications

Unspecified application

Species

Unspecified reactive species

Anne Cachat,Sébastien Alain Chevalier,Sandrine Alais,Nga Ling Ko,Lee Ratner,Chloé Journo,Hélène Dutartre,Renaud Mahieux
View all publications

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