Product: PKR Antibody
Catalog: AF6216
Description: Rabbit polyclonal antibody to PKR
Application: WB IHC IF/ICC
Reactivity: Human, Mouse
Mol.Wt.: 74kDa; 62kD(Calculated).
Uniprot: P19525
RRID: AB_2835097

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse
Clonality:
Polyclonal
Specificity:
PKR Antibody detects endogenous levels of total PKR.
RRID:
AB_2835097
Cite Format: Affinity Biosciences Cat# AF6216, RRID:AB_2835097.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Double stranded RNA activated protein kinase;; E2AK2_HUMAN; eIF-2A protein kinase 2; EIF2AK1; EIF2AK2; Eukaryotic translation initiation factor 2 alpha kinase 2; Eukaryotic translation initiation factor 2-alpha kinase 2; HGNC:9437; Interferon induced double stranded RNA activated protein kinase; Interferon inducible elF2 alpha kinase; Interferon inducible RNA dependent protein kinase; Interferon-induced, double-stranded RNA-activated protein kinase; Interferon-inducible RNA-dependent protein kinase; MGC126524; P1/eIF-2A protein kinase; P1/eIF2A protein kinase; p68 kinase; PKR; PPP1R83; PRKR; Protein kinase interferon inducible double stranded RNA dependent; Protein kinase RNA activated; Protein kinase RNA-activated; Protein phosphatase 1 regulatory subunit 83; Serine/threonine protein kinase TIK; Tyrosine protein kinase EIF2AK2;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P19525 E2AK2_HUMAN:

Highly expressed in thymus, spleen and bone marrow compared to non-hematopoietic tissues such as small intestine, liver, or kidney tissues. Colocalizes with GSK3B and TAU in the Alzheimer disease (AD) brain. Elevated levels seen in breast and colon carcinomas, and which correlates with tumor progression and invasiveness or risk of progression.

Description:
PKR a protein kinase of the PEK family. Upon binding double-stranded RNA, it becomes autophosphorylated and activated. Phosphorylates and inhibits the alpha subunit of eIF2 alpha, which leads to an inhibition of the initiation of protein synthesis.
Sequence:
MAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQVIIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKAVSPLLLTTTNSSEGLSMGNYIGLINRIAQKKRLTVNYEQCASGVHGPEGFHYKCKMGQKEYSIGTGSTKQEAKQLAAKLAYLQILSEETSVKSDYLSSGSFATTCESQSNSLVTSTLASESSSEGDFSADTSEINSNSDSLNSSSLLMNGLRNNQRKAKRSLAPRFDLPDMKETKYTVDKRFGMDFKEIELIGSGGFGQVFKAKHRIDGKTYVIKRVKYNNEKAEREVKALAKLDHVNIVHYNGCWDGFDYDPETSDDSLESSDYDPENSKNSSRSKTKCLFIQMEFCDKGTLEQWIEKRRGEKLDKVLALELFEQITKGVDYIHSKKLIHRDLKPSNIFLVDTKQVKIGDFGLVTSLKNDGKRTRSKGTLRYMSPEQISSQDYGKEVDLYALGLILAELLHVCDTAFETSKFFTDLRDGIISDIFDKKEKTLLQKLLSKKPEDRPNTSEILRTLTVWKKSPEKNERHTC

PTMs - P19525 As Substrate

Site PTM Type Enzyme
A2 Acetylation
S6 Phosphorylation
K20 Ubiquitination
S33 Phosphorylation
K61 Methylation
K69 Methylation
K69 Ubiquitination
K77 Ubiquitination
K80 Ubiquitination
S83 Phosphorylation P19525 (EIF2AK2)
T88 Phosphorylation P19525 (EIF2AK2)
T89 Phosphorylation P19525 (EIF2AK2)
T90 Phosphorylation P19525 (EIF2AK2)
S92 Phosphorylation
S93 Phosphorylation
Y101 Phosphorylation P19525 (EIF2AK2)
K140 Ubiquitination
Y142 Phosphorylation
K150 Ubiquitination
K159 Ubiquitination
Y162 Phosphorylation P19525 (EIF2AK2)
S179 Phosphorylation
S181 Phosphorylation
S242 Phosphorylation P19525 (EIF2AK2)
K253 Ubiquitination
T255 Phosphorylation P19525 (EIF2AK2)
Y257 Phosphorylation
T258 Phosphorylation P19525 (EIF2AK2)
K268 Ubiquitination
Y293 Phosphorylation P19525 (EIF2AK2)
K299 Ubiquitination
K304 Ubiquitination
K380 Ubiquitination
K385 Ubiquitination
K388 Ubiquitination
K400 Ubiquitination
S407 Phosphorylation
K408 Ubiquitination
K416 Ubiquitination
K426 Ubiquitination
K429 Ubiquitination
T437 Phosphorylation
S438 Phosphorylation
K440 Ubiquitination
K444 Ubiquitination
T446 Phosphorylation P19525 (EIF2AK2)
T451 Phosphorylation P51812 (RPS6KA3) , P28482 (MAPK1) , P19525 (EIF2AK2) , Q16539 (MAPK14)
Y454 Phosphorylation
S456 Phosphorylation
K509 Ubiquitination
K512 Ubiquitination
K517 Ubiquitination
K521 Ubiquitination
K522 Ubiquitination
R534 Methylation
T535 Phosphorylation
S542 Phosphorylation

PTMs - P19525 As Enzyme

Substrate Site Source
P04637-1 (TP53) S392 Uniprot
P05198 (EIF2S1) S49 Uniprot
P05198 (EIF2S1) S52 Uniprot
P06493 (CDK1) Y4 Uniprot
P10636-8 (MAPT) S262 Uniprot
P10636-8 (MAPT) S356 Uniprot
P19525 (EIF2AK2) S83 Uniprot
P19525 (EIF2AK2) T88 Uniprot
P19525 (EIF2AK2) T89 Uniprot
P19525 (EIF2AK2) T90 Uniprot
P19525 (EIF2AK2) Y101 Uniprot
P19525 (EIF2AK2) Y162 Uniprot
P19525 (EIF2AK2) S242 Uniprot
P19525 (EIF2AK2) T255 Uniprot
P19525 (EIF2AK2) T258 Uniprot
P19525 (EIF2AK2) Y293 Uniprot
P19525 (EIF2AK2) T446 Uniprot
P19525 (EIF2AK2) T451 Uniprot
P25963 (NFKBIA) S32 Uniprot
P42224 (STAT1) S727 Uniprot
Q15172 (PPP2R5A) S28 Uniprot

Research Backgrounds

Function:

IFN-induced dsRNA-dependent serine/threonine-protein kinase which plays a key role in the innate immune response to viral infection and is also involved in the regulation of signal transduction, apoptosis, cell proliferation and differentiation. 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). Inhibits viral replication via phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (EIF2S1), this phosphorylation impairs the recycling of EIF2S1 between successive rounds of initiation leading to inhibition of translation which eventually results in shutdown of cellular and viral protein synthesis. Also phosphorylates other substrates including p53/TP53, PPP2R5A, DHX9, ILF3, IRS1 and the HHV-1 viral protein US11. 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 proteosomal degradation. 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 proinflammatory cytokines and IFNs. 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. Can act as both a positive and negative regulator of the insulin signaling pathway (ISP). 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). Can regulate NLRP3 inflammasome assembly and the activation of NLRP3, NLRP1, AIM2 and NLRC4 inflammasomes. Can trigger apoptosis via FADD-mediated activation of CASP8. Plays a role in the regulation of the cytoskeleton by binding to gelsolin (GSN), sequestering the protein in an inactive conformation away from actin.

PTMs:

Autophosphorylated on several Ser, Thr and Tyr residues. Autophosphorylation of Thr-451 is dependent on Thr-446 and is stimulated by dsRNA binding and dimerization. Autophosphorylation apparently leads to the activation of the kinase. Tyrosine autophosphorylation is essential for efficient dsRNA-binding, dimerization, and kinase activation.

Subcellular Location:

Cytoplasm. Nucleus. Cytoplasm>Perinuclear region.
Note: Nuclear localization is elevated in acute leukemia, myelodysplastic syndrome (MDS), melanoma, breast, colon, prostate and lung cancer patient samples or cell lines as well as neurocytes from advanced Creutzfeldt-Jakob disease patients.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Highly expressed in thymus, spleen and bone marrow compared to non-hematopoietic tissues such as small intestine, liver, or kidney tissues. Colocalizes with GSK3B and TAU in the Alzheimer disease (AD) brain. Elevated levels seen in breast and colon carcinomas, and which correlates with tumor progression and invasiveness or risk of progression.

Subunit Structure:

Homodimer. Interacts with STRBP (By similarity). Interacts with DNAJC3. Forms a complex with FANCA, FANCC, FANCG and HSP70. Interacts with ADAR/ADAR1. Interacts with IRS1 (By similarity). The inactive form interacts with NCK1 and GSN. Interacts (via the kinase catalytic domain) with STAT3 (via SH2 domain), TRAF2 (C-terminus), TRAF5 (C-terminus) and TRAF6 (C-terminus). Interacts with MAP2K6, IKBKB/IKKB, NPM1, TARBP2, NLRP1, NLRP3, NLRC4 and AIM2. Interacts (via DRBM 1 domain) with DUS2L (via DRBM domain). Interacts with DHX9 (via N-terminus) and this interaction is dependent upon activation of the kinase.

(Microbial infection) Interacts with human herpes simplex virus 1 (HHV-1) protein US11 in an RNA-dependent manner.

(Microbial infection) The inactive form interacts with Toscana virus (TOS) NSS.

(Microbial infection) Interacts with herpes virus 8 protein v-IRF2; this interaction inhibits EIF2AK2 activation.

(Microbial infection) Interacts with vaccinia protein E3.

Family&Domains:

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum.   (View pathway)

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

References

1). Albendazole negatively regulates keratinocyte proliferation. CLINICAL SCIENCE, 2020 (PubMed: 32236445) [IF=6.0]

2). Risk subtyping and prognostic assessment of prostate cancer based on consensus genes. Communications Biology, 2022 (PubMed: 35293897) [IF=5.9]

Application: WB    Species: Human    Sample: C4-2 and PC-3 cells

Fig. 6 ITGA11 inhibits prostate cancer cell migration and invasion through PI3K/AKT signaling. a Overlapping coexpressed genes of ITGA11 in five cohorts and KEGG pathway enrichment analysis. b Correlation between ITGA11 expression and the activation level of PI3K/AKT signaling assessed by the HALLMARK PI3K/AKT/mTOR signaling signature. c Correlation between ITGA11 expression and the activation level of PI3K/AKT signaling assessed by the REACTOME PI3K/AKT activation signature. d Western blot analysis validated the inhibition of Ser473 p-AKT1/2/3 activation via knockdown of shITGA11 in both C4-2 and PC-3 cells. e Quantification of the western blotting results showing in d. Data are presented as the mean ± SD based on three independent experiments (ns not significant, *P < 0.05, **P < 0.01 by one-way ANOVA), bars represent mean values, error bars represent SD.

3). Targeted inhibition of CX3CL1 limits podocytes ferroptosis to ameliorate cisplatin-induced acute kidney injury. Molecular medicine (Cambridge, Mass.), 2023 (PubMed: 37875838) [IF=5.7]

Application: WB    Species: Mouse    Sample: kidney tissues

Fig. 6 CX3CL1 deficiency inhibited cisplatin-induced ER stress and HIF1A/HO-1 signal activation in kidney tissues. A Western blot analysis of proteins in kidney tissues that are associated with ER stress. B Western blotting was conducted to evaluate the HIF1A and HO-1 protein expression patterns in renal tissues. C Representative micrographs of CHOP and nephrin staining in renal tissues. Scale bar = 10 μm. D Representative micrographs of HO-1 and nephrin staining in renal tissues. Scale bar = 10 μm. E Western blot analysis was conducted to analyze the podocyte expression profiles of proteins associated with ER stress. F Podocytes’ HIF1A and HO-1 expression profiles were evaluated by western blotting. G Representative micrographs of podocytes stained with eIF2α and p-eIF2α. Scale bar = 10 μm. H Representative micrographs of podocytes stained with HIF1A and HO-1. Scale bar = 10 μm. (ER, endoplasmic reticulum; HIF1A/HO-1, hypoxia inducible factor 1-alpha/heme oxygenase-1; CHOP, CCAAT enhancer binding protein-homologous protein; eIF2α, eukaryotic translation initiation factor 2A; P value was calculated by one-way analysis of variance and Tukey’s test.

4). Knockdown of EIF2AK2-OAS1 axis reduces ATP production inducing AMPK phosphorylation to inhibit the malignant behavior of gastric cancer cells. Journal of bioenergetics and biomembranes, 2024 (PubMed: 38825632) [IF=3.0]

5). Molecular subtyping and prognostic assessment of prostate cancer based on consensus genes. Research Square, 2021

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