Recombinant
RabMAb

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

Rabbit recombinant monoclonal EIF2S1 (phospho S51) antibody [E90]. Validated in WB, IHC, Flow Cyt, Dot, ICC/IF and tested in Mouse, Rat, Human, Drosophila melanogaster, Plants and more.

Reviews (20)Q&A (2)References (117)

Overview

  • Product name

    Anti-EIF2S1 (phospho S51) antibody [E90]
    See all EIF2S1 primary antibodies

  • Description

    Rabbit monoclonal [E90] to EIF2S1 (phospho S51)

  • Host species

    Rabbit

  • Specificity

    Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157) only detects eIF-2a phosphorylated on Serine 51.

  • Tested applications

    Suitable 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

    corresponding to Human EIF2S1 (phospho S51).
    Database link: P05198
    (Peptide available as ab199382)

  • Positive control

    • IHC-P: Human liver carcinoma, breast carcinoma, cervical carcinoma, colon adenocarcinoma and hepatocellular carcinoma tissues. ICC/IF: Human epithelial cells WB: HeLa cells treated with Clyculin A and phosphatase whole cell lysate; PC-12 cell lysate Flow Cyt: HeLa cells Dot Blot: Antigen peptide.

  • General notes

      

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

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 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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 notes
    Is unsuitable for IP.

    • Target

    • 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 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 similarities

      Belongs 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 localization

      Cytoplasmic 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

    Images

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

      Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysates
      Lane 2 : HeLa treated with Calyculin A whole cell lysates
      Lane 3 : HeLa treated with Calyculin A and phosphatase whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : 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

    • Immunocytochemistry/ Immunofluorescence - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Immunocytochemistry/ Immunofluorescence - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)This image is courtesy of an Abreview submitted by Mr mike lindquist

      Paraformaldehyde-fixed human epithelial cells labeling EIF2SI using ab32157 at 1/100 dilution in ICC/IF, followed by an Alexa Fluor®647 conjugated goat anti-rabbit IgG.

      The cells were blocked with BSA before incubation with the antibody for 1 hour. 

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

      Immunohistochemical analysis of paraffin-embedded human liver carcinoma using ab32157 at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Flow Cytometry - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Flow Cytometry - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      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.
    • Dot Blot - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Dot Blot - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      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.
    • Western blot - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Western blot - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      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)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

      ab32157 showing positive staining in Breast carcinoma tissue at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

      ab32157 showing positive staining in Cervical carcinoma tissue at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

      ab32157 showing positive staining in Colonic adenocarcinoma tissue at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

      ab32157 showing positive staining in Hepatocellular carcinoma tissue at 1/50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Yadav AK  et al. AZD1208, a Pan-Pim Kinase Inhibitor, Has Anti-Growth Effect on 93T449 Human Liposarcoma Cells via Control of the Expression and Phosphorylation of Pim-3, mTOR, 4EBP-1, S6, STAT-3 and AMPK. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30654529) »
    • Cagnetta R  et al. Noncanonical Modulation of the eIF2 Pathway Controls an Increase in Local Translation during Neural Wiring. Mol Cell 73:474-489.e5 (2019). Read more (PubMed: 30595434) »
    See all 117 Publications for this product

    Publishing research using ab32157? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 22 Abreviews or Q&A

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Xenopus laevis Tissue lysate - whole (tedpole's head)
    Gel Running Conditions
    Non-reduced Denaturing (12%)
    Loading amount
    8 µg
    Treatment
    10uM octylphenol for 96hrs
    Specification
    tedpole's head
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Mr. Kyung noh Yoon

    Verified customer

    Submitted May 09 2019

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing (8 %)
    Loading amount
    20 µg
    Treatment
    Thapsigargin (0.5 uM)
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Mr. Jin Ik Kim

    Verified customer

    Submitted Apr 20 2019

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Pichia pastoris Cell lysate - whole cell (Pichia Pastoris GS115 (his-))
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    4.4 µg
    Specification
    Pichia Pastoris GS115 (his-)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 07 2018

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rabbit Cell lysate - whole cell (RK13 cells)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    200000 cells
    Specification
    RK13 cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 25 2017

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Brain Slices)
    Gel Running Conditions
    Reduced Denaturing (12% acrilamide)
    Loading amount
    20 µg
    Treatment
    2 ´g/ml for 4 hours
    Specification
    Brain Slices
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More

    Miss. Berta Anuncibay

    Verified customer

    Submitted May 04 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (MDA-MB-231 tumor cells grown on glass slides)
    Permeabilization
    Yes - 0.25% Triton-X100
    Specification
    MDA-MB-231 tumor cells grown on glass slides
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 14 2017

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (CLBL1 lymphoma cell line)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    10000 cells
    Specification
    CLBL1 lymphoma cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 13 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 13 2016

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 19 2016

    Western blot abreview for Anti-EIF2S1 (phospho S51) antibody [E90]

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 12 2016

    1-10 of 22 Abreviews or Q&A

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