PKR Antibody

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ProSci
Product Code: PSI-3947
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-3947-0.02mg0.02mg£150.00
Quantity:
PSI-3947-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation of PKR in Human Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: PKR 3947 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: PKR 3947 (1 μg/mL), PKR 3949 (1 μg/mL, and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 3 Immunofluorescence Validation of PKR in Mouse Lung</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse lung tissue labeling PKR with 3947 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
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<strong>Figure 4 Immunohistochemistry Validation of PKR in Rat Lung </strong><br> Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-PKR antibody (3947) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 5 Immunofluorescence Validation of PKR in Mouse Lung</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse lung tissue labeling PKR with 3947 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
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<strong>Figure 6 Immunohistochemistry Validation of PKR in Mouse Lung </strong><br> Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-PKR antibody (3947) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 1 Western Blot Validation of PKR in Human Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: PKR 3947 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: PKR 3947 (1 μg/mL), PKR 3949 (1 μg/mL, and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Immunofluorescence Validation of PKR in Mouse Lung</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse lung tissue labeling PKR with 3947 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 4 Immunohistochemistry Validation of PKR in Rat Lung </strong><br> Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-PKR antibody (3947) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunofluorescence Validation of PKR in Mouse Lung</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse lung tissue labeling PKR with 3947 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
<strong>Figure 6 Immunohistochemistry Validation of PKR in Mouse Lung </strong><br> Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-PKR antibody (3947) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Documents

Further Information

Additional Names:
PKR Antibody: PKR, PRKR, EIF2AK1, PKR, Interferon-induced, double-stranded RNA-activated protein kinase, Eukaryotic translation initiation factor 2-alpha kinase 2, eIF-2A protein kinase 2
Application Note:
WB: 1 μg/mL; IF: 20 ?g/mL; IHC-P: 2.5-5 ?g/mL.

Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in mouse and rat samples and Immunofluorescence in mouse samples. All other applications and species not yet tested.
Background:
PKR Antibody: The interferon-inducible, double-stranded RNA (dsRNA)-dependent protein kinase PKR is a member of the eukaryotic initiation factor-2 alpha (eIF2-alpha) kinase family, possessing serine-threonine kinase activity and two dsRNA-binding motifs that acts as part of the innate immune system. Upon binding dsRNA, PKR undergoes a conformational change leading to its activation and its phosphorylation of the translation factor eIF2, resulting in a general shutdown of protein synthesis and induction of apoptosis through upregulation of caspase-8 and capsase-9 activity in order to prevent the production of more viruses. To evade the antiviral effects of PKR, viruses have evolved multiple mechanisms, such as the inhibition of PKR by the non-structural protein (NS1) of the influenza virus. More recently, PKR has been implicated in several neurodegenerative diseases including Alzheimer, Huntington, and amyotrophic lateral sclerosis.
Background References:
  • Meurs et al. Cell 1990; 62:379-90.
  • Saelens et al. J. Biol. Chem. 2001; 276:41620-8.
  • Gil and Esteban. Apoptosis 2000; 5:107-14.
  • Gil et al. FEBS Lett. 2002; 19:3665-74.
Buffer:
PKR Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Immunogen:
Anti-PKR antibody (3947) was raised against a 14 amino acid synthetic peptide near the carboxy terminus of human PKR.

The immunogen is located within the last 50 amino acids of PKR.
ISOFORMS:
Human PKR has 2 isoforms, including isoform 1 (551aa, 62.1kD) and isoform S (510aa, 57.4kD. This antibody detects both human isoforms. Mouse PKR has 1 isoform (515aa, 58.3kD). Rat PKR has only one isoform identified so far (513aa, 58.3kD).
NCBI Gene ID #:
5610
NCBI Official Name:
eukaryotic translation initiation factor 2-alpha kinase 2
NCBI Official Symbol:
EIF2AK2
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 62kD

Observed: 68kD
Protein Accession #:
AAP57628
Protein GI Number:
31747519
Purification:
PKR Antibody is affinity chromatography purified via peptide column.
Research Area:
Innate Immunity
SPECIFICITY:
At least two different isoforms of PKR are known to exist; this antibody will detect both isoforms.
Swissprot #:
P19525
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Independent Antibody Validation in Cell lines (Figure 2) shows similar PKR expression profile in human and mouse cell lines detected by two independent anti-PKR antibodies that recognize different epitopes, 3947 against C-terminus domain and 3949 against N-terminus domain.  PKR proteins are detected in all the tested human cell lines except SK-N-SH cell line at different expression levels  by the two independent antibodies.  

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