Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

Overview

  • Product nameAnti-EIF2S1 (phospho S51) antibody [E90]
    See all EIF2S1 primary antibodies
  • Description
    Rabbit monoclonal [E90] to EIF2S1 (phospho S51)
  • SpecificityThe antibody only detects eIF-2a phosphorylated on Serine 51.
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cyt, Dot blot, IHC-Frmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Drosophila melanogaster, Plants, African Green Monkey, Neurospora crassa
    Predicted to work with: Cow, Pig
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human EIF2S1 (phospho S51).
    (Peptide available as ab199382)

  • Positive control
    • PC12 cell lysate, human liver carcinoma.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels.

    If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-EIF2S1 antibody (phospho S51) (Alexa Fluor® 488) [E90] (ab196460)

    Anti-EIF2S1 antibody (phospho S51) (Alexa Fluor® 647) [E90] (ab196191)

Properties

Applications

Our Abpromise guarantee covers the use of ab32157 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/250.
WB 1/500. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Dot blot 1/500.
IHC-Fr Use at an assay dependent concentration.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionFunctions 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. 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.
    • Sequence similaritiesBelongs to the eIF-2-alpha family.
      Contains 1 S1 motif domain.
    • Post-translational
      modifications
      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 (PubMed:15207627, PubMed:18032499). Phosphorylated; phosphorylation on Ser-52 by the EIF2AK4/GCN2 protein kinase occurs in response to amino acid starvation and UV irradiation.
    • Cellular localizationCytoplasmic granule. The cytoplasmic granules are stress granules which are a dense aggregation in the cytosol composed of proteins and RNAs that appear when the cell is under stress. Colocalizes with NANOS3 in the stress granules (By similarity).
    • Information by UniProt
    • Database links
      see all
    • Alternative names
      • EIF 2 alpha antibody
      • EIF 2 antibody
      • EIF 2A antibody
      • EIF 2alpha antibody
      • eIF-2-alpha antibody
      • eIF-2A antibody
      • EIF-2alpha antibody
      • EIF2 alpha antibody
      • EIF2 antibody
      • EIF2A antibody
      • EIF2S1 antibody
      • Eukaryotic translation initiation factor 2 subunit 1 alpha 35kDa antibody
      • Eukaryotic translation initiation factor 2 subunit 1 alpha antibody
      • Eukaryotic translation initiation factor 2 subunit 1 antibody
      • Eukaryotic translation initiation factor 2 subunit alpha antibody
      • IF2A_HUMAN antibody
      see all

    Anti-EIF2S1 (phospho S51) antibody [E90] images

    • All lanes : Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157) at 1/20000 dilution

      Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysates 20µg
      Lane 2 : HeLa (human cervix adenocarcinoma) treated with Calyculin A whole cell lysates 20µg
      Lane 3 : HeLa (human cervix adenocarcinoma) treated with Calyculin A and phosphatase whole cell lysates 20µg.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 36 kDa
      Observed band size : 36 kDa

      Blocking buffer and concentration: 2% BSA/TBST, Diluting buffer and concentration: 2% BSA /TBST, Exposure time: 10 seconds

    • Overlay histogram showing HeLa cells stained with ab32157 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32157, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157) at 1/500 dilution + PC12 cell lysate

      Predicted band size : 36 kDa
      Observed band size : 36 kDa

      Secondary antibody - anti-rabbit HRP (ab6721)

    • Immunohistochemical analysis of paraffin-embedded human liver carcinoma using ab32157 at 1/50 dilution.
    • Dot blot analysis on antigen peptide. A nitrocellulose membrane was spotted with (1) phospho-peptide and (2) non-phospho-peptide at 5, 1, and 0.1 ng, and then blotted with ab32157 at 1:500 dilution.
    • ab32157 at 1/100 staining human epithelial cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with BSA before incubation with the antibody for 1 hour. An Alexa-Fluor ® 647 conjugated goat anti-rabbit IgG was used as the secondary.

      See Abreview

    • ab32157 showing positive staining in Breast carcinoma tissue.

    • ab32157 showing positive staining in Cervical carcinoma tissue.

    • ab32157 showing positive staining in Colonic adenocarcinoma tissue.

    • ab32157 showing positive staining in Hepatocellular carcinoma tissue.

    References for Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

    This product has been referenced in:
    • Backlund M  et al. Endoplasmic reticulum stress increases AT1R mRNA expression via TIA-1-dependent mechanism. Nucleic Acids Res 44:3095-104 (2016). WB ; Human . Read more (PubMed: 26681690) »
    • Wengrod J  et al. Phosphorylation of eIF2a triggered by mTORC1 inhibition and PP6C activation is required for autophagy and is aberrant in PP6C-mutated melanoma. Sci Signal 8:ra27 (2015). WB ; Human . Read more (PubMed: 25759478) »

    See all 19 Publications for this product

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Sample Mouse Cell (B16 cells)
    Permeabilization Yes - 0.25% TritonX-100
    Specification B16 cells
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Sep 13 2016

    Application Western blot
    Sample Human Cell lysate - whole cell (MDA231 cells)
    Gel Running Conditions Reduced Denaturing (4-20% Tris Glycin gel)
    Loading amount 800000 cells
    Specification MDA231 cells
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Aug 19 2016

    Application Western blot
    Sample Human Cell lysate - whole cell (lymphocytes)
    Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
    Loading amount 15 µg
    Treatment 4ug/mL Tunicamycin or DMSO for 2 or 8 hours
    Specification lymphocytes
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 12 2016

    Application Western blot
    Sample Saccharomyces cerevisiae Cell lysate - whole cell (BY4741)
    Gel Running Conditions Reduced Denaturing (10%)
    Loading amount 10 µg
    Specification BY4741
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Dec 04 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate ph 7
    Sample Rat Tissue sections (rat brain)
    Specification rat brain
    Permeabilization No
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Aug 31 2014

    Application Western blot
    Loading amount 25 µg
    Gel Running Conditions Reduced Denaturing
    Sample Plants Tissue lysate - whole (Maize (Zea mays) endosperm)
    Specification Maize (Zea mays) endosperm
    Blocking step BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 24 2013


    The concentration of ab32157 (http://www.abcam.com/ab32157) lot GR26846-6is 0.574 mg/mL.

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Tissue lysate - whole (Liver)
    Loading amount 60 µg
    Specification Liver
    Gel Running Conditions Reduced Denaturing (gel 10%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Abcam user community

    Verified customer

    Submitted Jan 24 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (HepG2 cell line)
    Loading amount 30 µg
    Specification HepG2 cell line
    Treatment Asparaginase 1U/ml for 9h
    Gel Running Conditions Reduced Denaturing (gel 10%)
    Blocking step blocking solution (Roche as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Username

    Dr. Martina Chiu

    Verified customer

    Submitted Jan 17 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Neurospora crassa Cell lysate - whole cell (WT-74-OR23-1VA)
    Loading amount 200 µg
    Specification WT-74-OR23-1VA
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Gencer Sancar

    Verified customer

    Submitted Jul 14 2011

    1-10 of 15 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"