Product: Phospho-PERK (Thr982) Antibody
Catalog: DF7576
Description: Rabbit polyclonal antibody to Phospho-PERK (Thr982)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 125~180kDa; 125kD(Calculated).
Uniprot: Q9NZJ5
RRID: AB_2833024

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Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200, IF/ICC 1:100-500
*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,Rat
Prediction:
Pig(90%), Bovine(90%), Horse(100%), Sheep(90%), Rabbit(90%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-PERK (Thr982) Antibody detects endogenous levels of PERK only when phosphorylated at Thr982.
RRID:
AB_2833024
Cite Format: Affinity Biosciences Cat# DF7576, RRID:AB_2833024.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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

DKFZp781H1925; E2AK3_HUMAN; EC 2.7.11.1; Eif2ak3; Eukaryotic translation initiation factor 2 alpha kinase 3; Eukaryotic translation initiation factor 2-alpha kinase 3; Heme regulated EIF2 alpha kinase; HRI; HsPEK; Pancreatic eIF2 alpha kinase; Pancreatic eIF2-alpha kinase; PEK; PRKR like endoplasmic reticulum kinase; PRKR-like endoplasmic reticulum kinase; WRS;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q9NZJ5 E2AK3_HUMAN:

Ubiquitous. A high level expression is seen in secretory tissues.

Sequence:
MERAISPGLLVRALLLLLLLLGLAARTVAAGRARGLPAPTAEAAFGLGAAAAPTSATRVPAAGAVAAAEVTVEDAEALPAAAGEQEPRGPEPDDETELRPRGRSLVIISTLDGRIAALDPENHGKKQWDLDVGSGSLVSSSLSKPEVFGNKMIIPSLDGALFQWDQDRESMETVPFTVESLLESSYKFGDDVVLVGGKSLTTYGLSAYSGKVRYICSALGCRQWDSDEMEQEEDILLLQRTQKTVRAVGPRSGNEKWNFSVGHFELRYIPDMETRAGFIESTFKPNENTEESKIISDVEEQEAAIMDIVIKVSVADWKVMAFSKKGGHLEWEYQFCTPIASAWLLKDGKVIPISLFDDTSYTSNDDVLEDEEDIVEAARGATENSVYLGMYRGQLYLQSSVRISEKFPSSPKALESVTNENAIIPLPTIKWKPLIHSPSRTPVLVGSDEFDKCLSNDKFSHEEYSNGALSILQYPYDNGYYLPYYKRERNKRSTQITVRFLDNPHYNKNIRKKDPVLLLHWWKEIVATILFCIIATTFIVRRLFHPHPHRQRKESETQCQTENKYDSVSGEANDSSWNDIKNSGYISRYLTDFEPIQCLGRGGFGVVFEAKNKVDDCNYAIKRIRLPNRELAREKVMREVKALAKLEHPGIVRYFNAWLEAPPEKWQEKMDEIWLKDESTDWPLSSPSPMDAPSVKIRRMDPFATKEHIEIIAPSPQRSRSFSVGISCDQTSSSESQFSPLEFSGMDHEDISESVDAAYNLQDSCLTDCDVEDGTMDGNDEGHSFELCPSEASPYVRSRERTSSSIVFEDSGCDNASSKEEPKTNRLHIGNHCANKLTAFKPTSSKSSSEATLSISPPRPTTLSLDLTKNTTEKLQPSSPKVYLYIQMQLCRKENLKDWMNGRCTIEERERSVCLHIFLQIAEAVEFLHSKGLMHRDLKPSNIFFTMDDVVKVGDFGLVTAMDQDEEEQTVLTPMPAYARHTGQVGTKLYMSPEQIHGNSYSHKVDIFSLGLILFELLYPFSTQMERVRTLTDVRNLKFPPLFTQKYPCEYVMVQDMLSPSPMERPEAINIIENAVFEDLDFPGKTVLRQRSRSLSSSGTKHSRQSNNSHSPLPSN

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
100
Dog
100
Xenopus
100
Chicken
100
Pig
90
Bovine
90
Sheep
90
Rabbit
90
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9NZJ5 As Substrate

Site PTM Type Enzyme
T177 Phosphorylation
Y268 Phosphorylation
T441 Phosphorylation
S447 Phosphorylation
K452 Ubiquitination
S455 Phosphorylation
Y464 Phosphorylation
Y474 Phosphorylation
Y480 Phosphorylation
Y481 Phosphorylation
Y484 Phosphorylation
Y485 Phosphorylation
S555 Phosphorylation
T557 Phosphorylation
T561 Phosphorylation
S567 Phosphorylation
K581 Ubiquitination
Y585 Phosphorylation
Y619 Phosphorylation Q9NZJ5 (EIF2AK3)
K622 Ubiquitination
K669 Ubiquitination
S688 Phosphorylation
S694 Phosphorylation
T705 Phosphorylation
S715 Phosphorylation
S719 Phosphorylation
T802 Phosphorylation P31751 (AKT2) , P31749 (AKT1)
S803 Phosphorylation
S804 Phosphorylation
S811 Phosphorylation
S844 Phosphorylation
S845 Phosphorylation
S854 Phosphorylation
S856 Phosphorylation
T861 Phosphorylation
T862 Phosphorylation
T982 Phosphorylation
S1094 Phosphorylation
S1096 Phosphorylation
S1109 Phosphorylation
S1111 Phosphorylation

PTMs - Q9NZJ5 As Enzyme

Substrate Site Source
P05198 (EIF2S1) S49 Uniprot
P05198 (EIF2S1) S52 Uniprot
Q9BY44 (EIF2A) S265 Uniprot
Q9NZJ5 (EIF2AK3) Y619 Uniprot

Research Backgrounds

Function:

Metabolic-stress sensing protein kinase that phosphorylates the alpha subunit of eukaryotic translation initiation factor 2 (eIF-2-alpha/EIF2S1) on 'Ser-52' during the unfolded protein response (UPR) and in response to low amino acid availability. Converts phosphorylated eIF-2-alpha/EIF2S1 either in a global protein synthesis inhibitor, leading to a reduced overall utilization of amino acids, or to a translation initiation activator of specific mRNAs, such as the transcriptional activator ATF4, and hence allowing ATF4-mediated reprogramming of amino acid biosynthetic gene expression to alleviate nutrient depletion. Serves as a critical effector of unfolded protein response (UPR)-induced G1 growth arrest due to the loss of cyclin-D1 (CCND1). Involved in control of mitochondrial morphology and function.

PTMs:

Oligomerization of the N-terminal ER luminal domain by ER stress promotes PERK trans-autophosphorylation of the C-terminal cytoplasmic kinase domain at multiple residues including Thr-982 on the kinase activation loop (By similarity). Autophosphorylated. Phosphorylated at Tyr-619 following endoplasmic reticulum stress, leading to activate its tyrosine-protein kinase activity. Dephosphorylated by PTPN1/TP1B, leading to inactivate its enzyme activity.

N-glycosylated.

ADP-ribosylated by PARP16 upon ER stress, which increases kinase activity.

Subcellular Location:

Endoplasmic reticulum membrane>Single-pass type I membrane protein.

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

Ubiquitous. A high level expression is seen in secretory tissues.

Subunit Structure:

Forms dimers with HSPA5/BIP in resting cells (By similarity). Oligomerizes in ER-stressed cells (By similarity). Interacts with DNAJC3 and MFN2 (By similarity). Interacts with TMEM33. Interacts with PDIA6.

Family&Domains:

The lumenal domain senses perturbations in protein folding in the ER, probably through reversible interaction with HSPA5/BIP.

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

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

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

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

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· 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 > Herpes simplex infection.

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

References

1). Targeting Mitochondria-Located circRNA SCAR Alleviates NASH via Reducing mROS Output. CELL (PubMed: 32931733) [IF=64.5]

Application: WB    Species: human    Sample: fibroblasts

(J) Representative western blots for CHOP, p-PERK, ATF4, and p-eIF2a in normal fibroblasts treated with palmitate (n = 3 patients).

2). VAPB-mediated ER-targeting stabilizes IRS-1 signalosomes to regulate insulin/IGF signaling. Cell Discovery (PubMed: 37528084) [IF=33.5]

3). Boost therapy of hepatocellular carcinoma by amplifying vicious cycle between mitochondrial oxidative stress and endoplasmic reticulum stress via biodegradable ultrasmall nanoparticles and old drug. Nano Today [IF=17.4]

4). Dual Inhibition of Endoplasmic Reticulum Stress and Oxidation Stress Manipulates the Polarization of Macrophages under Hypoxia to Sensitize Immunotherapy. ACS Nano (PubMed: 34414762) [IF=17.1]

5). A non-canonical pathway regulates ER stress signaling and blocks ER stress-induced apoptosis and heart failure. Nature Communications (PubMed: 28743963) [IF=16.6]

Application: WB    Species: rat    Sample:

Fig. 4 AGGF1 protein therapy regulates ER stress signaling and apoptosis. TAC or sham mice were treated with AGGF1 or PBS (left) and characterized (n = 6/group, **P < 0.01). a AGGF1 regulates TAC-induced ER stress signaling in mice. Protein extracts from heart samples were used for western blot analysis for ER stress signaling markers (n = 6/group, **P < 0.01). b Representative images for immunostaining analysis of heart sections for KDEL receptor-positive cells. Scale bar, 50 μm. c Representative images of TUNEL staining for apoptosis from heart sections. Scale bar, 50 μm. d Western blot analysis for apoptosis markers in heart tissues (n = 6/group, **P < 0.01). e Real-time RT-PCR analyses for ATF4, ATF6, CHOP, Ero1α, and GADD34 in heart tissues from TAC or sham mice treated with AGGF1 or PBS (n = 5/group, **P < 0.01). f Western blot analysis showing that AGGF1 protein treatment increased the levels of ATF4 and p-eIF2α, and decreased the level of sXBP1 in H9C2 cells treated with ISO for 48 h. The effects of AGGF1 were abolished by overexpression of CHOP by transient transfection of an expression plasmid as compared with the empty vector. No effect was observed for p-PERK (n = 3/group, **P < 0.01, N.S., Non-significant). g Real-time RT-PCR analysis for GADD34 in H9C2 cells transfected with an expression plasmid for CHOP or empty vector control, and then treated with ISO in combination with AGGF1 or PBS for 48 h (n = 3/group, *P < 0.05). Data are shown as the mean ± s.d. from at least three independent experiments. For a–e, statistical analysis was carried out by a Student’s two-tailed t-test; for f, g, statistical analysis was carried out by one-way analysis of variance

6). Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. NUCLEIC ACIDS RESEARCH (PubMed: 32710621) [IF=14.9]

Application: WB    Species: mouse    Sample: liver

Figure 7. Effect of A22 on ameliorating apoptosis, ER stress, inflammation, metabolic syndrome, and fibrogenesis in HF diet-fed mice. (A) Effect of A22 on BCL-2 gene transcription. (B) Effect of A22 on BAX gene transcription. (C) Effect of A22 on expressions of apoptosis-related proteins in liver. The extracted proteins from the liver were immunoblotted with specific antibodies, and quantified based on the loading control of ACTIN. (D) Effect of A22 on ER stress. The UPR proteins (IRE-1, PERK, elF-2 and CHOP) were analyzed by using western Blot. (E) Effect of A22 on expressions of inflammatory factors. (F) Effect of A22 on expressions of fibrogenic proteins.

7). Targeted regulation of lymphocytic ER stress response with an overall immunosuppression to alleviate allograft rejection. Biomaterials (PubMed: 33798960) [IF=14.0]

Application: WB    Species: mouse    Sample: CD8+T cells

Fig. 4.|The phenotype and quantity of T-8 under UPR intervention.(K-L) Immunoblot analysis of XBP-1s, p-PERK and CHOP expression in T-8 treated with different UPRi-loaded liposomes at indicated phases.

8). Induction of EnR stress by Melatonin enhances the cytotoxic effect of Lapatinib in HER2-positive breast cancer. Cancer Letters (PubMed: 34153400) [IF=9.7]

Application: WB    Species: Human    Sample: HER2-positive breast cancer cells

Fig. 4. The combination of Melatonin and Lapatinib results in substantial EnR stress-induced UPR in HER2-positive breast cancer cells. (A) Gene Set Enrichment Analysis (GSEA) of UPR/ER stress pathway signatures in the Melatonin (2 mM) plus Lapatinib (1 μM) -treated HCC1954 cells versus Lapatinib alone treated cells. Drug treatment time: 24 h. NES, FDR q values and p values of the correlation was shown. (B) Heatmap showing the expression of UPR/ER stress pathway genes in the HCC1954 cells treated with Melatonin and Lapatinib as single-agents or in combination. Row Z-score value is shown. (C) Quantitative RT-PCR analysis of the expression of ATF4, ATF6, PERKA and IRE1 in HCC1954 and MDA-MB-453 cells subjected to drug treatment as indicated. ACTB was used as an endogenous control. HCC1954: 2 mM Melatonin, 1 μM Lapatinib; MDA-MB-453: 2 mM Melatonin, 2 μM Lapatinib. (D) Western Blot analysis of the protein levels in the HER2- positive breast cancer cells subjected to drug treatment as indicated. HCC1954 and MDA-MB-453 cells were treated as in (C); MCF7/HER2: 1 mM Melatonin, 1 μM Lapatinib. Vinculin was used as a loading control. (E–F) Cell viability was measured by crystal violet assay for the HCC1954 treated with Lapatinib at different concentrations and with or without BFA (0.1 μM). (E) 96-well format; (F) 24-well format. (G) Western Blot analysis of the protein levels in the HCC1954 cells treated with Lapatinib (1.0 μM) and BFA (0.1 μM), either alone or in combination. Data was shown as Mean ± S.D. *p < 0.05, **p < 0.01, ***p < 0.001 (Student’s t-test).

9). A marine-derived small molecule induces immunogenic cell death against triple-negative breast cancer through ER stress-CHOP pathway. International Journal of Biological Sciences (PubMed: 35541893) [IF=9.2]

Application: WB    Species: human    Sample: MDA-MB-231 cells

Figure 2. | MHO7 induced ER stress and cell cycle arrest in TNBC cells.(D) The expression of BiP/p-PERK/p-eIF2α/ATF4/CHOP were measured by western blot under MHO7 treatment in MDA-MB-231 cells.

10). Icariside II enhances cisplatin-induced apoptosis by promoting endoplasmic reticulum stress signalling in non-small cell lung cancer cells. International Journal of Biological Sciences (PubMed: 35342361) [IF=9.2]

Application: WB    Species: mouse    Sample: A549/DDP cells

Figure 5.| IS enhances cisplatin-induced apoptosis by promoting ER stress in A549/DDP cells.(H) IS promotes the expression of the proapoptotic transcription factor CHOP. Cells were harvested after 24 h of treatment. Data were from at least three independent experiments. 4-PBA: 4-phyenylbutyric acid.

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