eIF2 alpha Antibody | Affinity Biosciences

Product:	eIF2 alpha Antibody
Catalog:	AF6087
Description:	Rabbit polyclonal antibody to eIF2 alpha
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat, Pig
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	38kDa; 36kD(Calculated).
Uniprot:	P05198
RRID:	AB_2834856

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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.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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Related Downloads

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,Pig

Prediction:

Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)

Clonality:

Polyclonal

Specificity:

eIF2 alpha Antibody detects endogenous levels of total eIF2 alpha.

RRID:

AB_2834856
Cite Format: Affinity Biosciences Cat# AF6087, RRID:AB_2834856.

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

EIF 2 alpha; EIF 2; EIF 2A; EIF 2alpha; eIF-2-alpha; eIF-2A; EIF-2alpha; EIF2 alpha; EIF2; EIF2A; EIF2S1; Eukaryotic translation initiation factor 2 subunit 1 alpha 35kDa; Eukaryotic translation initiation factor 2 subunit 1 alpha; Eukaryotic translation initiation factor 2 subunit 1; Eukaryotic translation initiation factor 2 subunit alpha; IF2A_HUMAN;

Immunogens

Immunogen:

Uniprot:

P05198(IF2A_HUMAN) >>Visit HPA database.

Gene(ID):

EIF2S1(1965)

Description:

eIF2A a translation initiation factor that functions in the early steps of protein synthesis by forming a ternary complex with GTP and initiator tRNA. This complex binds to a 40s ribosomal subunit, followed by mRNA binding to form a 43S preinitiation complex.

Sequence:

P05198 IF2A_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MPGLSCRFYQHKFPEVEDVVMVNVRSIAEMGAYVSLLEYNNIEGMILLSELSRRRIRSINKLIRIGRNECVVVIRVDKEKGYIDLSKRRVSPEEAIKCEDKFTKSKTVYSILRHVAEVLEYTKDEQLESLFQRTAWVFDDKYKRPGYGAYDAFKHAVSDPSILDSLDLNEDEREVLINNINRRLTPQAVKIRADIEVACYGYEGIDAVKEALRAGLNCSTENMPIKINLIAPPRYVMTTTTLERTEGLSVLSQAMAVIKEKIEEKRGVFNVQMEPKVVTDTDETELARQMERLERENAEVDGDDDAEEMEAKAED

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 - P05198 As Substrate

P05198

Site	PTM Type	Enzyme	Source
S5	Phosphorylation		Uniprot
K12	Ubiquitination		Uniprot
S26	Phosphorylation		Uniprot
S49	Phosphorylation	Q9NZJ5 (EIF2AK3) , Q9BQI3 (EIF2AK1) , P19525 (EIF2AK2) , Q75MR0 (HRI)	Uniprot
S52	Phosphorylation	B4DS64 (PRKRIR) , P51812 (RPS6KA3) , P19525 (EIF2AK2) , P28482 (MAPK1) , Q75MR0 (HRI) , Q9BQI3 (EIF2AK1) , Q9NZJ5 (EIF2AK3) , Q05655 (PRKCD) , Q16539 (MAPK14) , Q9P2K8 (EIF2AK4)	Uniprot
S58	Phosphorylation		Uniprot
K61	Ubiquitination		Uniprot
K80	Ubiquitination		Uniprot
Y82	Phosphorylation		Uniprot
S86	Phosphorylation		Uniprot
K87	Ubiquitination		Uniprot
S91	Phosphorylation		Uniprot
K97	Ubiquitination		Uniprot
K101	Acetylation		Uniprot
K101	Ubiquitination		Uniprot
K104	Ubiquitination		Uniprot
K106	Ubiquitination		Uniprot
S110	Phosphorylation		Uniprot
K123	Ubiquitination		Uniprot
R133	Methylation		Uniprot
K141	Acetylation		Uniprot
K141	Ubiquitination		Uniprot
K143	Acetylation		Uniprot
K143	Ubiquitination		Uniprot
Y147	Phosphorylation		Uniprot
Y150	Phosphorylation		Uniprot
K154	Methylation		Uniprot
K154	Ubiquitination		Uniprot
S158	Phosphorylation		Uniprot
T185	Phosphorylation		Uniprot
K190	Ubiquitination		Uniprot
Y200	Phosphorylation		Uniprot
Y202	Phosphorylation		Uniprot
K209	Ubiquitination		Uniprot
S219	Phosphorylation		Uniprot
K226	Ubiquitination		Uniprot
Y235	Phosphorylation		Uniprot
T245	Phosphorylation		Uniprot
K259	Ubiquitination		Uniprot
K265	Ubiquitination		Uniprot
R266	Methylation		Uniprot
K276	Ubiquitination		Uniprot
T279	Phosphorylation		Uniprot
T281	Phosphorylation		Uniprot
T284	Phosphorylation		Uniprot
K312	Ubiquitination		Uniprot

Research Backgrounds

P05198_IF2A_HUMAN

Function:

Functions in the early steps of protein synthesis by forming a ternary complex with GTP and initiator tRNA. This complex binds to a 40S ribosomal subunit, followed by mRNA binding to form a 43S pre-initiation complex. Junction of the 60S ribosomal subunit to form the 80S initiation complex is preceded by hydrolysis of the GTP bound to eIF-2 and release of an eIF-2-GDP binary complex. In order for eIF-2 to recycle and catalyze another round of initiation, the GDP bound to eIF-2 must exchange with GTP by way of a reaction catalyzed by eIF-2B.

PTMs:

Substrate for at least 4 kinases: EIF2AK1/HRI, EIF2AK2/PKR, EIF2AK3/PERK and EIF2AK4/GCN2. Phosphorylation stabilizes the eIF-2/GDP/eIF-2B complex and prevents GDP/GTP exchange reaction, thus impairing the recycling of eIF-2 between successive rounds of initiation and leading to global inhibition of translation. Phosphorylated; phosphorylation on Ser-52 by the EIF2AK4/GCN2 protein kinase occurs in response to amino acid starvation and UV irradiation (By similarity).

Subcellular Location:

Cytoplasm>Stress granule.
Note: Colocalizes with NANOS3 in the stress granules.

Subunit Structure:

Heterotrimer composed of an alpha, a beta and a gamma chain. Component of an EIF2 complex at least composed of CELF1/CUGBP1, CALR, CALR3, EIF2S1, EIF2S2, HSP90B1 and HSPA5. Interaction with METAP2 protects EIF2S1 from inhibitory phosphorylation (By similarity). Interacts with ABCF1 isoform 2. Associates with ribosomes. Interacts with DDX3X in an RNA-independent manner.

Family&Domains:

Belongs to the eIF-2-alpha family.

Research Fields

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

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

· Genetic Information Processing > Translation > RNA transport.

· 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 > 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.

References

1). 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.

2). Betulinic acid chemosensitizes breast cancer by triggering ER stress-mediated apoptosis by directly targeting GRP78. Cell Death & Disease (PubMed: 29802332) [IF=9.0]

Application: WB Species: human Sample: MCF-7,MDA-MB-231

Fig. 5 BA triggers breast cancer cells apoptosis via ER stress-mediated pathway. a MCF-7 and MDA-MB-231 cells were treated with the indicated concentrations of BA for 24 h, and the protein levels of ER stress-associated signals were stimulated by BA in a dose-dependent manner, including GRP78, p-PERK/PERK, p-eIF2α/eIF2α, CHOP, and caspase-12. b MCF-7 and MDA-MB-231 cells were treated with BA alone or in combination with taxol for 24 h, the expression levels of GRP78, p-PERK/PERK, p-eIF2α/eIF2α, CHOP, and caspase-12 were also significantly upregulated following drug administration, especially in the co-treatment group, indicating the ER stress-mediated apoptosis pathway was aggravatedly activated by drug combination.

3). -bambuterol and its enantiomers: Potential improvement of (R)-bambuterol in mice with colitis. International Immunopharmacology (PubMed: 34974400) [IF=5.6]

4). 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 (PubMed: 31632274) [IF=5.6]

Application: WB Species: rat Sample: liver

FIGURE 5 | PA treatment attenuated HFD-induced VLDLR expression in rats. (A) Representative immunoreactive bands of eIF2α, p-eIF2α, ATF4, and VLDLR

5). CTRP1 attenuates cerebral ischemia/reperfusion injury via the PERK signaling pathway. Frontiers in Cell and Developmental Biology (PubMed: 34422821) [IF=5.5]

Application: WB Species: Rat Sample: cortex

FIGURE 7 The function of CTRP1 on apoptosis and PERK signal pathway was inhibited by CCT020312. (A) Apoptosis was analyzed by TUNEL, n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm. (B) Western blot analyzed the expression CTRP1, BAX, CHOP, GRP78, ATF4, p-PERK, PERK, p-eIF2α, and eIF2α in the cortex. n = 6 per group. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 vs. sham group, ####p < 0.0001, ##p < 0.01, #p < 0.05 vs. MCAO/R + LV-NC group, @@p < 0.01, @p < 0.05 vs. MCAO/R + LV-CTRP1 group.

6). ATP citrate lyase inhibitor triggers endoplasmic reticulum stress to induce hepatocellular carcinoma cell apoptosis via p‐eIF2α/ATF4/CHOP axis. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (PubMed: 33393219) [IF=5.3]

Application: WB Species: Human Sample: HepG2 cells

FIGURE 5 ACLY inhibitor triggers ER stress and activates p‐eIF2α/ATF4/CHOP axis in vitro. Western blot analysis of (A) ER stress‐related proteins (p‐eIF2α, eIF2α, ATF4 and CHOP) and (B) UPR signal transduction molecules (p‐PERK, PERK, p‐IRE1α, IRE1α and sXBP1) in HepG2 cells after administration of BMS‐303141. ATF4p‐eIF2α, eIF2α were activated 3 h post‐treatment; CHOP was activated 8 h post‐treatment. (* P < .05, ** P < .01 and *** P < .001, compared with control group) (C) Western blot analysis of protein expression after ATF4 knockdown. (D) Annexin V‐FITC/PI double staining was performed to determine the apoptosis rate of HepG2 cells after ATF4 knockdown via flow cytometry. (* P < .05, ** P < .01 and *** P < .001, compared with con siRNA group). All experiments were repeated 3 times

7). MiR-185-5p ameliorates endoplasmic reticulum stress and renal fibrosis by downregulation of ATF6. LABORATORY INVESTIGATION (PubMed: 32514126) [IF=5.0]

Application: WB Species: human Sample: HK2 cells

Supplementary figure 2. |miR-185-5p suppresses the activation of the IRE1/XBP1 and PERK/eIF2α/ATF4 branches in TGF-β1-induced HK2 cells.(C, D) Western blot analysis of p-PERK, PERK, p-eIF2α, eIF2α, and ATF4 proteins.Data are presented as the mean ± SD (n = 3) calculated from one-way ANOVA with Tukey’s test.

8). EndophilinA2 protects against angiotensin II-induced cardiac hypertrophy by inhibiting angiotensin II type 1 receptor trafficking in neonatal rat cardiomyocytes. JOURNAL OF CELLULAR BIOCHEMISTRY (PubMed: 29923351) [IF=4.0]

Application: WB Species: rat Sample: cardiomyocytes

FIGURE 4| EndoA2 modulated the MAPK signaling pathway in response to ERS. G-L, Western blotting results show the expression levels of GRP78, p-PERK, p-eiF2α, ATF4, and CHOP after EndoA2 siRNA knockdown. Densitometric analyses show that EndoA2 siRNA knockdown increased the expression levels of GRP78, p-PERK, p-eiF2α, ATF4, and CHOP compared with those in the Ang II group. Transfection with neg had no significant effects (n = 5, *P < 0.05 vs control, #P < 0.05 vs AngII).

9). Supplementation of branched-chain amino acids to a reduced-protein diet improves growth performance in piglets: involvement of increased feed intake and direct muscle growth-promoting effect. BRITISH JOURNAL OF NUTRITION (PubMed: 27079773) [IF=3.6]

Application: WB Species: pig Sample:

Fig. 3. Abundance of phosphorylated and total eukaryotic translation initiation factor 2α (eIF2α) protein in the hypothalamus of piglets fed positive control (PC, ) diet, reduced-protein negative control (NC, ) diet or two NC diets supplemented with a 1- or 2-fold dose of BCAA (test 1 ( ) and test 2 ( ), respectively) in Expt 1. Western blotting method was used. Values were normalised using β-actin or relative total protein as an internal control. Data are expressed relative to expressions in the PC group; values are means (n 4–6/ group), with their standard errors represented by vertical bars. a,b Mean values with unlike letters were significantly different (P <0·05).

10). Inonotus obliquus extract alleviates myocardial ischemia/reperfusion injury by suppressing endoplasmic reticulum stress. Molecular Medicine Reports (PubMed: 33236154) [IF=3.4]

Application: WB Species: rat Sample: myocardium

Figure 6.| Expression of SIRT1, GRP78, p‑PERK, p‑eIF2α, CHOP, caspase‑12 and the mRNA levels of caspase‑12 in the myocardium. (A) Representative blots of SIRT1, GRP78, p‑PERK, p‑eIF2α, CHOP and caspase‑12. Semiquantitative analysis of (B) SIRT1, (C) GRP78, (D) p‑PERK, (E) p‑eIF2α, (F) CHOP and (G) caspase‑12.

Application: WB Species: Rat Sample: myocardium

Figure 6. Expression of SIRT1, GRP78, p-PERK, p-eIF2α, CHOP, caspase-12 and the mRNA levels of caspase-12 in the myocardium. (A) Representative blots of SIRT1, GRP78, p-PERK, p-eIF2α, CHOP and caspase-12. Semiquantitative analysis of (B) SIRT1, (C) GRP78, (D) p-PERK, (E) p-eIF2α, (F) CHOP and (G) caspase-12. (H) The mRNA levels of caspase-12 in the myocardium. Data are presented as the mean ± standard deviation. n=3. ##P<0.01 vs. Sham. *P<0.05 and **P<0.01 vs. MI/R. IOE, Inonotus obliquus extract; SIRT1, NAD-dependent protein deacetylase sirtuin-1; GRP78, glucose-regulated protein 78; PERK, protein kinase R-like endoplasmic reticulum kinase; eIF2α, eukaryotic translation initiation factor 2 subunit α; CHOP, C/EBP homologous protein; p, phosphorylated; MI/R, myocardial ischemia/reperfusion.

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eIF2 alpha Antibody - #AF6087