Phospho-eIF2 alpha (Ser51)[Ser52] Antibody

Product:	Phospho-eIF2 alpha (Ser51)[Ser52] Antibody
Catalog:	AF3087
Description:	Rabbit polyclonal antibody to Phospho-eIF2 alpha (Ser51)[Ser52]
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	38kDa; 36kD(Calculated).
Uniprot:	P05198
RRID:	AB_2834524

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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 1:50-1:500, 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:

Phospho-eIF2 alpha (Ser51) Antibody detects endogenous levels of eIF2 alpha only when phosphorylated at Ser52, which site historically referenced as Ser51.

RRID:

AB_2834524
Cite Format: Affinity Biosciences Cat# AF3087, RRID:AB_2834524.

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

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, 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). A marine-derived small molecule induces immunogenic cell death against triple-negative breast cancer through ER stress-CHOP pathway. International Journal of Biological Sciences, 2023 (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.

3). Betulinic acid chemosensitizes breast cancer by triggering ER stress-mediated apoptosis by directly targeting GRP78. Cell Death & Disease, 2018 (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.

4). Modulating endoplasmic reticulum stress in APP/PS1 mice by Gomisin B and Osthole in Bushen-Yizhi formula: Synergistic effects and therapeutic implications for Alzheimer's disease. Phytomedicine, 2023 (PubMed: 37586159) [IF=7.9]

5). Up-regulation of thioredoxin system by puerarin inhibits lipid uptake in macrophages. Free radical biology & medicine, 2021 (PubMed: 33242606) [IF=7.4]

6). Formononetin attenuates cigarette smoke-induced COPD in mice by suppressing inflammation, endoplasmic reticulum stress, and apoptosis in bronchial epithelial cells via AhR/CYP1A1 and AKT/mTOR signaling pathways. Phytotherapy research : PTR, 2024 (PubMed: 38191199) [IF=7.2]

7). Microcystin-leucine arginine induced the apoptosis of GnRH neurons by activating the endoplasmic reticulum stress resulting in a decrease of serum testosterone level in mice. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY, 2021 (PubMed: 33396074) [IF=6.8]

Application: WB Species: Mice Sample: GT1–7 cells

Fig. 4. Activation of the endoplasmic reticulum stress (ERs) in GnRH neurons and GT1–7 cells after MC-LR treatment. (A) MC-LR triggered ultrastructural changes of ERs in hypothalamus neurons. Drinking water including 15 μg/L MCLR was fed to mice for 180 days without interruption, and the ultrastructure of ERs (black arrows) in control and MC-LR treatment group were detected by electron microscope. (B) Increase of intracellular Ca2+ concentration in GT1–7 cells following exposure to MC-LR. Fluo-3/AM fluorescent probes were used to measure levels of intracellular Ca2+ through immunofluorescence assay (scale bar = 100 µm). (C) The intensity of Fluo-3/AM was detected by image-pro plus (n = 3). (D) Mice were given drinking water comprising MC-LR for 180 sustained days. Expressions of GRP78, ATF6, ATF4, PERK, CHOP, eIF2α, peIF2α, and LC3B in hypothalamus tissues were assessed by Western blotting (n = 5 mice/ group). (E) GT1–7 cells were plated in 6-well plates at 2.0 × 105 cells per well. Cells were exposed to MC-LR for 24 h at different doses as indicated. Expressions of GRP78, ATF6, ATF4, PERK, CHOP, eIF2α, p- eIF2α, and LC3B in GT1–7 cells were assessed by Western blotting. (F) GT1–7 cells were plated in 6-well plates at 2.0 × 105 cells per well. Cells were pretreated with ERs inhibitor 4- Phenyl butyric acid (4- PBA) at 5 mM for 1 h, followed by the treatment with MC-LR (1000 nM) for 24 h. The expression levels of ERs related proteins and LC3B were measured.

8). Icariin improves the cognitive function of APP/PS1 mice via suppressing endoplasmic reticulum stress. LIFE SCIENCES, 2019 (PubMed: 31400352) [IF=6.1]

Application: WB Species: mouse Sample: hippocampus

Fig.6 |Effects of ICA on the PERK signaling in the hippocampus of APP/PS1 mice. The phosphorylation of PERK and eIF2α, as well ATF4 and CHOP proteins expression were increased in APP/PS1 mice, ICA treatment significantly decreased their levels. (A, B) Immunoblots of PERK, p-PERK, eIF2α, p-eIF2α, ATF4 and CHOP.

9). Cyanidin-3-O-glucoside and protocatechuic acid alleviate heat stress-induced testicular damage. Food & Function, 2023 (PubMed: 36756975) [IF=6.1]

10). Intranasal Administration of GRP78 Protein (HSPA5) Confers Neuroprotection in a Lactacystin-Induced Rat Model of Parkinson's Disease. International journal of molecular sciences, 2024 (PubMed: 38612761) [IF=5.6]

Application: WB Species: Rat Sample: nigral tissue

Figure 4 Exogenous GRP78 blocks the pro-apoptotic GRP78/eIF2α/CHOP/caspase-3,9 signaling pathway of the UPR in nigral tissue in a rat model of Parkinson’s disease. Nigral content of (a) GRP78, (b) phosphorylated to total eIF2α, (c) CHOP. (d) Representative immunoblots. Nigral content of (e) cleaved caspase-9, (f) cleaved caspase-3. (g) Representative immunoblots. Western blot analysis of the nigral tissue was conducted with the antibodies against GRP78 (1:1000, rabbit, Abcam, Cambridge, UK), eIF2a (1:750, rabbit, Affinity Biosciences, Zhenjiang, China), pSer51-eIF2a (1:1000, rabbit, Abcam, Cambridge, UK), CHOP (1:1000, rabbit, Affinity Biosciences, Zhenjiang, China), cleaved caspase-9 (1:1000, rabbit, Affinity Biosciences, Zhenjiang, China), and cleaved caspase-3 (1:1000, rabbit, Affinity Biosciences, Zhenjiang, China). Staining with anti-β-Actin antibodies (1:1000, mouse, Santa Cruz Biotechnology, Dallas, TX, USA) was used as the loading control. The results are presented as percentages of the control (panels (a–c,e,f)). Bar charts indicate mean values with standard errors. The dots, squares, triangles and rhombus indicate individual values per rat. Two-way ANOVA test followed by Tukey’s post hoc analysis were performed to determine the effects of GRP78 therapy. Asterisks indicate significant differences between groups according to Tukey’s post hoc tests

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Phospho-eIF2 alpha (Ser51)[Ser52] Antibody - #AF3087