Product: PERK Antibody
Catalog: AF5304
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 125~140kD; 125kD(Calculated).
Uniprot: Q9NZJ5
RRID: AB_2837789

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*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(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
PERK Antibody detects endogenous levels of total PERK.
RRID:
AB_2837789
Cite Format: Affinity Biosciences Cat# AF5304, RRID:AB_2837789.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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.

Description:
Phosphorylates the alpha subunit of eukaryotic translation-initiation factor 2 (EIF2), leading to its inactivation and thus to a rapid reduction of translational initiation and repression of global protein synthesis.
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
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Chicken
100
Rabbit
100
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). Li X et al. Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. Nucleic Acids Res 2020 Sep 4;48(15):8255-8268. (PubMed: 32710621) [IF=19.160]

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.

2). Zhang S et al. The role of ATF6 in Cr (VI)-induced apoptosis in DF-1 cells. J Hazard Mater 2020 Nov 19;124607. (PubMed: 33243643) [IF=14.224]

Application: WB    Species: chicken    Sample: DF-1 cells

Fig. 6. Effects of ATF-6 on the different apoptotic pathway. (A) The mRNA expression levels of PERK, ATF-6 and Caspase-12 as detected by RT-qPCR. (B) PERK, ATF- 6 and Caspase-12 detected by Western blot and quantitative analysis the protein level. (C) The mRNA expression levels of Caspase-9, Bcl-2 and Bax inhibitor-1 as detected by RT-qPCR. (D) Proteins expression of pro Caspase-9, Bcl-2 and Bax detected by Western blot, quantitative analysis the protein level. (E) The mRNA expression levels of Caspase-8 as detected by RT-qPCR. (F) Expression of Caspase-8 and quantitative analysis of the protein level. All data were expressed as relative values against their respective control group. Data were presented as means ± SD (n = 3). NS P > 0.05, *P < 0.05, **P < 0.01.

3). Mo C et al. Development of erianin-loaded dendritic mesoporous silica nanospheres with pro-apoptotic effects and enhanced topical delivery. J Nanobiotechnology 2020 Mar 30;18(1):55 (PubMed: 32228604) [IF=10.435]

Application: WB    Species: Human    Sample: HaCaT cells

Fig. 5 Efect of erianin and E/DMSNs on cytosolic calcium levels and ERS signaling pathway. a Flow cytometry analysis of cytosolic calcium levels in HaCaT cells after treatment with erianin and E/DMSNs for 24 h. b Relative MFI of control in (a) analyzed by fow cytometry. c The expressions of PERK, ATF6, IRE1α, and CHOP proteins after treatment with erianin and E/DMSNs for 24 h. d Quantitation of PERK, ATF6, IRE1α, and CHOP proteins normalized to β-actin in (c) by using Image J software. Values are represented as means±SD (n=3). *p<0.05, **p<0.01, ***p<0.005, ****p<0.001, signifcantly diferent compared with the erianin group. ##p<0.01, ###p<0.005, ####p<0.001, signifcantly diferent compared with the control group

Application: WB    Species: Human    Sample: HaCaT cells

Fig. 5 Effect of erianin and E/DMSNs on cytosolic calcium levels and ERS signaling pathway. a Flow cytometry analysis of cytosolic calcium levels in HaCaT cells after treatment with erianin and E/DMSNs for 24 h. b Relative MFI of control in (a) analyzed by flow cytometry. c The expressions of PERK, ATF6, IRE1α, and CHOP proteins after treatment with erianin and E/DMSNs for 24 h. d Quantitation of PERK, ATF6, IRE1α, and CHOP proteins normalized to β-actin in (c) by using Image J software. Values are represented as means ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001, significantly different compared with the erianin group. ##p < 0.01, ###p < 0.005, ####p < 0.001, significantly different compared with the control group

4). Mo C et al. Development of erianin-loaded dendritic mesoporous silica nanospheres with pro-apoptotic effects and enhanced topical delivery. J Nanobiotechnology 2020 Mar 30;18(1):55 (PubMed: 32228604) [IF=10.435]

5). Xiong S et al. AdipoRon Protects against Tubular Injury in Diabetic Nephropathy by Inhibiting Endoplasmic Reticulum Stress. Oxid Med Cell Longev 2020 Aug 6;2020:6104375. (PubMed: 32832003) [IF=7.310]

Application: WB    Species: mice    Sample: kidneys

Figure 3.(d) Representative immunoblots of AdipoR1, p-AMPK, and T-AMPK. β-Actin was used as a loading control. (e) Relative band intensity.

6). Zhao YS et al. Hydrogen and Oxygen Mixture to Improve Cardiac Dysfunction and Myocardial Pathological Changes Induced by Intermittent Hypoxia in Rats. Oxid Med Cell Longev 2019 Mar 7;2019:7415212 (PubMed: 30984338) [IF=7.310]

Application: WB    Species: rat    Sample: endoplasmic reticulum

Figure 3: |The H2-O2 mixture-induced inhibition of ER stress caused by CIH for 35 d: (a) the ER stress markers GRP 78, caspase 12, and CHOP protein expressions; (b–d) the ratios of p-PERK, p-eIF2α/eIF2α, and p-IRE 1/IRE 1 in the left ventricle

7). Tian JH et al. Zonisamide, an antiepileptic drug, alleviates diabetic cardiomyopathy by inhibiting endoplasmic reticulum stress. Acta Pharmacol Sin 2020 Jul 9. (PubMed: 32647341) [IF=7.169]

Application: WB    Species: mice    Sample: NRCMs

Fig. 6 Zonisamide alleviates HG-induced cardiac hypertrophy and apoptosis in cultured primary neonatal rat cardiomyocytes (NRCMs) via suppression of activated ER stress. NRCMs were pretreated with 5 mM 4-PBA (an inhibitor of ERS) or 10 ng/mL tunicamycin (Tm, an ERS inducer) for 2 h and then exposed to glucose (33 mM) in the presence or absence of ZNS (3 μM) for 24 h. a–b Representative and quantitative images showing the protein expression of ERS markers, including GRP78, XBP-1s, ATF6, p-PERK, PERK, ATF4, CHOP, and Hrd1. c Immunofluorescence staining of cardiomyocytes with phalloidin (red) and cell nuclei with DAPI (blue), Scale bar = 50 μm. d Quantitative analysis of cell surface area by ImageJ software. e–f Representative Western blotting and analysis of Bax and Bcl-2 expression. g–h Representative and quantitative images of GRP78, ATF6, p-PERK, PERK, ATF4, and CHOP expression. All values are the fold changes normalized to their control group. The results are presented as the means ± SEM (n = 6). *P < 0.05, **P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. HG; $P < 0.05, $$P < 0.01 vs. HG + ZNS.

8). Sun ZM et al. Resveratrol protects against CIH-induced myocardial injury by targeting Nrf2 and blocking NLRP3 inflammasome activation. Life Sci 2020 Jan 27:117362 (PubMed: 31996295) [IF=6.780]

Application: WB    Species: Rat    Sample: heart tissue

Fig. 4. Resveratrol treatment weakened cardiac ER stress in rats exposed to CIH. A. Representative western blot images related to ER stress. B. p-PERK/ERK ratio. C. p-IRE/IRE ratio. D. GRP78 expression. n = 6. *p < 0.05 vs. control group; #p < 0.05 vs. CIH group. △ There was a CIH × resveratrol interaction.

9). Wu L et al. Involvement of miR‐27a‐3p in diabetic nephropathy via affecting renal fibrosis, mitochondrial dysfunction, and endoplasmic reticulum stress. J Cell Physiol 2020 Jul 21. (PubMed: 32691413) [IF=6.513]

Application: WB    Species: mice    Sample: kidney

FIGURE 4 Effect of miR‐27a‐3p on endoplasmic reticulum stress and apoptosis in the renal cortex of db/db mice. (a) Western blot analysis of p‐IRE1α, IRE1α, XBP1s, p‐PERK, PERK, p‐eIF2α, eIF2α, and CHOP proteins in the kidney tissues of mice. (b) Apoptosis in renal cortex was visualized by TUNEL immunohistochemical staining. Data are presented as mean ± SD (n = 6). CHOP, C/EBP homologous protein; eIF2α, eukaryotic initiation factor 2 α; IRE1α, inositol‐requiring transmembrane kinase/endoribonuclease 1α; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; XBP1, X‐box binding protein 1

10). Shi W et al. Identification of dihydrotanshinone I as an ERp57 inhibitor with anti-breast cancer properties via the UPR pathway. Biochem Pharmacol 2021 May 29;190:114637. (PubMed: 34062127) [IF=6.100]

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

Fig. 5.| DHT activated ER stress and UPR signaling pathways. MDA-MB-231 cells were incubated with 3 μM DHT for the indicated time. (A) Western blot analysis was performed with antibodies against Sec61γ, BiP, p-PERK/PERK, p-eIF2α/eIF2α, and ATF4. The quantitative densitometric analysis of Sec61γ (B), BiP (C), p-PERK/PERK (D), p-eIF2α/eIF2α (E), and ATF4 (F) was carried out.

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