Phospho-EIF2S1 (Ser51) Monoclonal antibody

Phospho-EIF2S1 (Ser51) Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

5

CloneNo.

1A4A11

Cat no : 68023-1-Ig

Synonyms

EIF2S1, EIF2S1/EIF2A, EIF2, EIF 2alpha, EIF 2A

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Validation Data Gallery

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  WB analysis using 68023-1-Ig

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 68023-1-Ig (Phospho-EIF2S1 (Ser51) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin (HRP-66240) antibody as loading control.

• 

  WB analysis using 68023-1-Ig

  Non-treated and Calyculin A treated PC-3 cells were subjected to SDS PAGE followed by western blot with 68023-1-Ig (Phospho-EIF2S1 (Ser51) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin (HRP-66240) antibody as loading control.

• 

  WB analysis using 68023-1-Ig

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 68023-1-Ig (Phospho-EIF2S1 (Ser51) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin (HRP-66009) antibody as loading control.

• 

  WB analysis using 68023-1-Ig

  Non-treated and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 68023-1-Ig (Phospho-EIF2S1 (Ser51) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin (HRP-66240) antibody as loading control.

• 

  WB analysis using 68023-1-Ig

  Non-treated and Calyculin A treated HSC-T6 cells were subjected to SDS PAGE followed by western blot with 68023-1-Ig (Phospho-EIF2S1 (Ser51) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Tubulin (HRP-66240) antibody as loading control.

• 

  WB analysis of HEK-293 using 68023-1-Ig

  Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 68023-1-Ig (Phospho-EIF2S1 (Ser51) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  IF Staining of HeLa using 68023-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed Calyculin A treated HeLa cells using Phospho-EIF2S1 (Ser51) antibody (68023-1-Ig, Clone: 1A4A11 ) at dilution of 1:800 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM002).

• 

  FC experiment of PC-3 using 68023-1-Ig

  1X10^6 PC-3 cells untreated (dashed lines) or treated with Calyculin A (red) were intracellularly stained with 0.5 ug Anti-Human Phospho-EIF2S1 (Ser51) (68023-1-Ig, Clone:1A4A11) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000, or 0.5 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Tested Applications

• Positive WB detected in: PC-3 cells, HeLa cells, Calyculin A treated HEK-293 cells, NIH/3T3 cells, HSC-T6 cells, Calyculin A treated HeLa cells, Calyculin A treated PC-3 cells, Calyculin A treated NIH/3T3 cells, Calyculin A treated HSC-T6 cells
• Positive IF/ICC detected in: Calyculin A treated HeLa cells
• Positive FC (Intra) detected in: Calyculin A treated PC-3 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.50 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 5 publications below

Product Information

68023-1-Ig targets Phospho-EIF2S1 (Ser51) in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: eukaryotic translation initiation factor 2, subunit 1 alpha, 35kDa
• Calculated Molecular Weight: 36 kDa
• Observed Molecular Weight: 36 kDa
• GenBank Accession Number: NM_004094
• Gene Symbol: EIF2S1
• Gene ID (NCBI): 1965
• RRID: AB_2918767
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

EIF2S1 is one subunit of the translation initiation factor EIF2, which catalyzes the first regulated step of protein synthesis initiation, promoting the binding of the initiator tRNA to 40S ribosomal subunits. This complex binds to a 40S ribosomal subunit, followed by mRNA binding to form a 43S preinitiation 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. This phosphorylation stabilizes the eIF2-GDP-eIF2B complex and inhibits the turnover of eIF2B. Induction of PKR by IFN-γ and TNF-α induces potent phosphorylation of eIF2α at Ser51.

Publications

Species	Application	Title
human	WB	bioRxiv; Mitosis exit followed by death in interphase prevents the development of polyploid giant cancer cells; Authors - Juan Jesus Vicente
human	WB	Cell Death Dis; AURKA emerges as a vulnerable target for KEAP1-deficient non-small cell lung cancer by activation of asparagine synthesis; Authors - Bing Deng
human	WB	J Transl Med; The microtubule targeting agent ST-401 triggers cell death in interphase and prevents the formation of polyploid giant cancer cells; Authors - Juan Jesus Vicente
human	WB	Environ Pollut; Copper exposure induces mitochondrial dysfunction and hepatotoxicity via the induction of oxidative stress and PERK/ATF4 -mediated endoplasmic reticulum stress; Authors - Meng Li
human,mouse	WB	Microbiol Res; Berberine ameliorates vascular dysfunction by downregulating TMAO-endoplasmic reticulum stress pathway via gut microbiota in hypertension; Authors - Zhichao Wang