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Human PERK Antibody

R&D Systems, part of Bio-Techne | Catalog # AF3999

R&D Systems, part of Bio-Techne
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AF3999
AF3999-SP

Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant human PERK
Ala29-Gln230
Accession # Q9NZJ5

Specificity

Detects human PERK in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human PERK Antibody

Detection of Human PERK antibody by Western Blot.

Detection of Human PERK by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and NTera-2 human testicular embryonic carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Human PERK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3999) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PERK at approximately 130 kDa (as indicated). This experiment was conducted using Immunoblot Buffer Group 1.
Western Blot Shows Human PERK Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human PERK Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and PERK knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human PERK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3999) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for PERK at approximately 150 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human PERK by Western Blot

Detection of Human PERK by Western Blot

Activation of the ER stress signaling pathway leads to the LX-2 cell apoptosis induced by VP-16.(a) Western blotting analysis of ER stress-associated proteins in LX-2 cells after treatment with VP-16 for 72 h. (b) After exposure to VP-16 for 72 h, the protein phosphorylation levels of PERK and eIF2 alpha were analyzed by western blotting. (c) After exposure to VP-16 for 72 h, the proteins of the IRE1 alpha /ASK1/JNK signaling pathway were analyzed by western blotting. (d,e) LX-2 cells were pretreated with a JNK inhibitor (SP600125, 40 μM) for 1 h, and then treated with 4 μM VP-16 for 72 h. (d) Cell viability was measured using the CCK-8 assay. The percentage of apoptosis cells was analyzed by flow cytometry. The data are presented as the mean ± SD of at least three independent experiments performed in triplicates. **P < 0.01, compared with the group treated with VP-16 alone. (e) Western blotting analysis of the cell lysates was performed using the indicated antibodies. In all of the western blotting analyses, beta-actin was used as a loading control. *P < 0.05, compared with the control group. #P < 0.05, compared with the group treated with VP-16 alone. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27680712), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human PERK Antibody

Application
Recommended Usage

Knockout Validated

PERK is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in PERK knockout HeLa cell line.

Western Blot

1 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line and NTera-2 human testicular embryonic carcinoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using AF3999 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PERK

PERK, a type 1 ER membrane kinase, mediates eIF2 alpha phosphorylation at Ser51 during the UPR (unfolded protein response). Protein synthesis is inhibited, thereby reducing the burden of protein substrate for the ER folding and degradation mechanism. Phosphorylation of eIF2 alpha also selectively promotes the expression of UPR target genes such as Chop and BiP. PERK may also play a role in tumor cell adaptation to hypoxic stress by regulating the translation of angiogenic factors necessary for the development of functional microvessels. Mutations in PERK are responsible for the rare autosomal-recessive disorder, WRS (Wolcott-Rallison syndrome).

Long Name

Eukaryotic Translation Initiation Factor 4B

Alternate Names

EIF2AK3, PEK, WRS

Entrez Gene IDs

9451 (Human); 13666 (Mouse); 29702 (Rat)

Gene Symbol

EIF2AK3

UniProt

Additional PERK Products

Product Documents for Human PERK Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human PERK Antibody

For research use only

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