Name: Anti-PKR antibody [Y117]
SKU: ab32506
Rating: 4 (3 reviews)

Anti-PKR antibody [Y117] (ab32506) is a rabbit monoclonal antibody that is used to detect PKR in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human samples.

- Specificity confirmed with PKR knockout cell line validation

Related conjugates and formulations

ab207342
Conjugated
Alexa Fluor® 647 Anti-PKR antibody [Y117]
ab207341
Conjugated
Alexa Fluor® 488 Anti-PKR antibody [Y117]
ab305722
Conjugated
PE Anti-PKR antibody [Y117]
ab305724
Conjugated
HRP Anti-PKR antibody [Y117]
ab308756
Conjugated
Alkaline Phosphatase Anti-PKR antibody [Y117]
ab310471
Conjugated
Alexa Fluor® 594 Anti-PKR antibody [Y117]
ab312477
Conjugated
Alexa Fluor® 568 Anti-PKR antibody [Y117]
ab312001
Conjugated
Alexa Fluor® 555 Anti-PKR antibody [Y117]
ab239817
Carrier free
Anti-PKR antibody [Y117] - BSA and Azide free
ab305723
Conjugated
APC Anti-PKR antibody [Y117]
ab321606
Conjugated
Alexa Fluor® 750 Anti-PKR antibody [Y117]

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] (AB32506), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/80 - 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000 - 1/20000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes For unpurified use at 1/250 - 1/500

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

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Target data

See full target information EIF2AK2

Function

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).

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: Y117
Purification technique: Affinity purification Protein A
Specificity: This antibody does not cross-react with other GCN2 family members.
Concentration

Storage

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

Notes

What is this antibody validated in?

Anti-PKR antibody [Y117] (ab32506) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), in Human samples.

What is the molecular weight of PKR?

Anti-PKR [Y117] (ab32506) specifically detects a band for PKR (UniProt: P19525) at a molecular weight of 62kDa.

Recommended positive controls

WB: MCF-7, HEK293 and HeLa whole cell lysate (ab150035) and human liver carcinoma tissue lysate.ICC/IF: Wild type HAP1, MCF-7 and HeLa cells.IHC-P: Human liver carcinoma and colon carcinoma tissue.

Trusted by the scientific community

Anti-PKR [Y117] (ab32506) was first used in a scientific publication in 2006 and has been cited over 20 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed

The specificity of Anti-PKR antibody [Y117] (ab32506) has been confirmed by Western blot testing in EIF2AK2 Knockout HAP1 cells.

Other related products

We have a range of other formats of antibody clone [Y117] also available for your convenience:
ab32506, Alexa Fluor® 488 - ab207341, Alexa Fluor® 647 - ab207342, Carrier free - ab239817, PE - ab305722, APC - ab305723, HRP - ab305724, Alkaline Phosphatase - ab308756, Alexa Fluor® 594 - ab310471, Alexa Fluor® 555 - ab312001, Alexa Fluor® 568 - ab312477, Alexa Fluor® 750 - ab321606

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] (ab32506)
ab32506 staining PKR in human liver carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
Western blot - Anti-PKR antibody [Y117] (ab32506)
Lanes 1, 3 and 5: PKR knockout HAP1 cell lysate (20 μg)
Lanes 2, 4 and 6: Wild-type HAP1 cell lysate (20 μg)
Lanes 1 and 2: Green signal from target - ab32506 observed at 62 kDa
Lanes 3 and 4: Red signal from loading control - Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa
Lanes 5 and 6: Merged (red and green) signal
ab32506 was shown to specifically react with PKR when PKR knockout samples were used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32506 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-PKR antibody [Y117] (ab32506)
Predicted band size: 62 kDa
Western blot - Anti-PKR antibody [Y117] (ab32506)
Purified format.
All lanes: Western blot - Anti-PKR antibody [Y117] (ab32506) at 1/20000 dilution
Lane 1: MCF-7 (human breast carcinoma) whole cell lysates at 20 µg
Lane 2: HEK293 (human embryonic kidney) whole cell lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 62 kDa
Observed band size: 68 kDa
Immunoprecipitation - Anti-PKR antibody [Y117] (ab32506)
ab32506 immunoprecipitating PKR. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at a dilution of 1/10000.
Lane 1: HEK293 (human embryonic kidney) whole cell lysate (10ug)
Lane 2: HEK293 (human embryonic kidney) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32506 in HEK293 (human embryonic kidney) whole cell lysate
All lanes: Immunoprecipitation - Anti-PKR antibody [Y117] (ab32506)
Predicted band size: 62 kDa
Observed band size: 40 kDa, 68 kDa
Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] (ab32506)
ab32506 staining PKR in wild-type HAP1 cells (top panel) and PKR knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32506 at 1/400 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.
This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 4% formaldehyde (10 min).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] (ab32506)
ab32506 staining PKR in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 were used as counterstains for primary antibody ab32506 and secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 respectively and DAPI was used as a nuclear counterstain.
Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120)
Negative control 2: Mouse primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077)
Western blot - Anti-PKR antibody [Y117] (ab32506)
All lanes: Western blot - Anti-PKR antibody [Y117] (ab32506) at 1/10000 dilution
All lanes: MCF-7 cell lysate
Predicted band size: 62 kDa
Observed band size: 68 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] (ab32506)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using unpurified ab32506 at 1/100 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] (ab32506)
Immunofluorescent staining of HeLa cells using unpurified ab32506 at 1/250 dilution.
Flow Cytometry (Intracellular) - Anti-PKR antibody [Y117] (ab32506)
Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling PKR with purified ab32506 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-PKR antibody [Y117] (ab32506)
PKR western blot using anti-PKR antibody [Y117] ab32506. Publication image and figure legend from Liu, G. & Zhang, W., 2018, Braz J Med Biol Res, PubMed 29898032.

ab32506 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab32506 please see the product overview.
Effect of HOTAIR on PKR expression; A, qRT-PCRrevealed overexpression of HOTAIR promoted the expression of PKR;B, Western blot analysis revealedoverexpression of HOTAIR promoted the expression of PKR. Data arereported as means±SD. *P<0.05; **P<0.01 (ANOVA). HOTAIR: HOXantisense intergenic RNA; PKR: RNA-dependent protein kinase; qRT-PCR:quantitative real-time polymerase chain reaction.