PERK Antibody | Affinity Biosciences

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

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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100ul	$280	In stock
200ul	$350	In stock

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MS Report

HPLC Report

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*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(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.

Conjugate:

Unconjugated.

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:

Q9NZJ5(E2AK3_HUMAN) >>Visit HPA database.

Gene(ID):

EIF2AK3(9451)

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:

Q9NZJ5 E2AK3_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

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

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9NZJ5 As Substrate

Q9NZJ5

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

PTMs - Q9NZJ5 As Enzyme

Q9NZJ5

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

Research Backgrounds

Q9NZJ5_E2AK3_HUMAN

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.

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). Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. NUCLEIC ACIDS RESEARCH, 2020 (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.

2). The role of ATF6 in Cr (VI)-induced apoptosis in DF-1 cells. JOURNAL OF HAZARDOUS MATERIALS, 2021 (PubMed: 33243643) [IF=13.6]

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). Development of erianin-loaded dendritic mesoporous silica nanospheres with pro-apoptotic effects and enhanced topical delivery. JOURNAL OF NANOBIOTECHNOLOGY, 2020 (PubMed: 32228604) [IF=10.2]

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). Zonisamide, an antiepileptic drug, alleviates diabetic cardiomyopathy by inhibiting endoplasmic reticulum stress. Acta Pharmacologica Sinica, 2020 (PubMed: 32647341) [IF=8.2]

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.

5). Mitofusin2 Ameliorated Endoplasmic Reticulum Stress and Mitochondrial Reactive Oxygen Species Through Maintaining Mitochondria-Associated Endoplasmic Reticulum Membrane Integrity in Cisplatin-Induced Acute Kidney Injury. Antioxidants & Redox Signaling, 2023 (PubMed: 37053105) [IF=6.6]

6). Pterostilbene Prevents Tunicamycin-Induced Intestinal Barrier Damage by Targeting Endoplasmic Reticulum Stress, Oxidative Stress, Autophagy, and Gut Microbiota. Journal of Agricultural and Food Chemistry, 2022 (PubMed: 36225099) [IF=6.1]

7). Resveratrol protects against CIH-induced myocardial injury by targeting Nrf2 and blocking NLRP3 inflammasome activation. LIFE SCIENCES, 2020 (PubMed: 31996295) [IF=6.1]

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.

8). Network-based identification and mechanism exploration of active ingredients against Alzheimer's disease via targeting endoplasmic reticulum stress from traditional chinese medicine. Computational and structural biotechnology journal, 2024 (PubMed: 38261917) [IF=6.0]

9). Identification of dihydrotanshinone I as an ERp57 inhibitor with anti-breast cancer properties via the UPR pathway. BIOCHEMICAL PHARMACOLOGY, 2021 (PubMed: 34062127) [IF=5.8]

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.

10). Protective Effect of Patchouli Alcohol Against High-Fat Diet Induced Hepatic Steatosis by Alleviating Endoplasmic Reticulum Stress and Regulating VLDL Metabolism in Rats. Frontiers in Pharmacology, 2019 (PubMed: 31632274) [IF=5.6]

Application: WB Species: rat Sample: liver

FIGURE 4 | PA treatment attenuated HFD-induced ER stress in rats. (A) Representative immunoreactive bands of GRP78, PERK, p-PERK, IRE1α, p-IRE1α, and ATF6

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PERK Antibody - #AF5304