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<strong>Figure 1 Western Blot Validation of PKR in MCF-7 Cells </strong><br>Loading: 15 μg of lysates per lane.Antibodies: PKR 3949 (A: 1 μg/mL; B: 2 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 Western Blot Validation of PKR in Human Cell Lines </strong><br>Loading: 15 μg of lysates per lane.Antibodies: PKR 3949 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of PKR in Rat Lung </strong><br>Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-PKR antibody (3949) 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.
Figure 1 Western Blot Validation of PKR in MCF-7 Cells
Loading: 15 μg of lysates per lane.Antibodies: PKR 3949 (A: 1 μg/mL; B: 2 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
Trial Size

PKR Antibody

CATALOG NUMBER: 3949

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Mouse (86%)
Immunogen:
Anti-PKR antibody (3949) was raised against a 14 amino acid synthetic peptide near the amino terminus of human PKR.
The immunogen is located within the first 50 amino acids of PKR.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB
Application Note:
WB: 1-2 μg/mL; IHC-P: 5 μg/mL.
Antibody validated: Western Blot in human and mouse samples and Immunohistochemistry in rat samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1208 - THP-1 cell lysate
Positive Control 2:
Cat. No. 1462 - Rat Lung Tissue Lysate
Positive Control 3:
Cat. No. 1224 - Daudi Cell Lysate
Positive Control 4:
Cat. No. 1202– A431 Cell Lysate
Predicted Molecular Weight:
Predicted: 62kD
Observed: 68kD
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.  

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).
Purification:
PKR Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
PKR Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
PKR antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
EIF2AK2
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
Protein Accession Number:
NP_002750
Protein Gi Number:
31747519
Ncbi Gene Id Number:
5610
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
Meurs et al. Cell 1990; 62:379-90.
Background Reference 2:
Saelens et al. J. Biol. Chem. 2001; 276:41620-8.
Background Reference 3:
Gil and Esteban. Apoptosis 2000; 5:107-14.
Background Reference 4:
Gil et al. FEBS Lett. 2002; 19:3665-74.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

3949

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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