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Phospho-EIF2S1 (S51) Recombinant Rabbit Monoclonal Antibody [SZ01-06]

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Catalog# ET1603-14

Phospho-EIF2S1 (S51) Recombinant Rabbit Monoclonal Antibody [SZ01-06]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-EIF2S1 (S51) Recombinant Rabbit Monoclonal Antibody [SZ01-06]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser51 of Human eIF-2a.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Molecular Weight

Predicted band size: 36 kDa

Positive Control

HeLa treated with 50nM Calyculin A for 3 hours whole cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate, C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate, THP-1 cell lysate, C2C12 cell lysate, mouse spleen tissue, rat spleen tissue, human liver tissue, human pancreas tissue, mouse brain tissue, mouse placenta tissue, mouse pancreas tissue, human prostate carcinoma tissue, human breast carcinoma tissue, human colon carcinoma tissue, Hela.

Conjugation

unconjugated

Clone Number

SZ01-06

RRID

AB_3069669

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:500

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

Phosphorylation of the eukaryotic initiation factor 2 (eIF2) α subunit is a well-documented mechanism to downregulate protein synthesis under a variety of stress conditions. Eukaryotic initiation factor 2 binds GTP and Met-tRNAi and transfers Met-tRNA to the 40S subunit to form the 43S preinitiation complex. eIF2 promotes a new round of translation initiation by exchanging GDP for GTP, a reaction catalyzed by eIF2B. Kinases that are activated by viral infection (PKR), endoplasmic reticulum stress (PERK/PEK), amino acid deprivation (GCN2), or heme deficiency (HRI) can phosphorylate the α subunit of eIF2. 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.

Background References

1. Montalbano R et al. Endoplasmic reticulum stress plays a pivotal role in cell death mediated by the pan-deacetylase inhibitor panobinostat in human hepatocellular cancer cells. Transl Oncol 6:143-57 (2013).

2. Kanai R et al. Effect of 34.5 deletions on oncolytic herpes simplex virus activity in brain tumors. J Virol 86:4420-31 (2012).

Sequence Similarity

Belongs to the WD repeat EIF2A family.

Tissue Specificity

Widely expressed. Expressed at higher level in pancreas, heart, brain and placenta.

Subcellular Location

Stress granule, Cytoplasm.

UNIPROT

SwissProt: P05198 Human

SwissProt: Q6ZWX6 Mouse

SwissProt: P68101 Rat

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-EIF2S1 (S51) on different lysates with Rabbit anti-Phospho-EIF2S1 (S51) antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa whole cell lysate (15 µg/Lane)<br />Lane 2: HeLa treated with 50nM Calyculin A for 3 hours whole cell lysate (15 µg/Lane)<br />Lane 3: NIH/3T3 whole cell lysate (15 µg/Lane)<br />Lane 4: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate (15 µg/Lane)<br />Lane 5: C6 whole cell lysate (20 µg/Lane)<br />Lane 6: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate (20 µg/Lane)<br /><br />Predicted band size: 36 kDa<br />Observed band size: 36 kDa<br /><br />Exposure time: Lane 1-4: 2 minutes; Lane 5-6: 23 seconds；<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-EIF2S1 (S51) on different lysates with Rabbit anti-Phospho-EIF2S1 (S51) antibody (ET1603-14) at 1/2,000 dilution.

    Lane 1: HeLa whole cell lysate (15 µg/Lane)
    Lane 2: HeLa treated with 50nM Calyculin A for 3 hours whole cell lysate (15 µg/Lane)
    Lane 3: NIH/3T3 whole cell lysate (15 µg/Lane)
    Lane 4: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate (15 µg/Lane)
    Lane 5: C6 whole cell lysate (20 µg/Lane)
    Lane 6: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate (20 µg/Lane)

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

    Exposure time: Lane 1-4: 2 minutes; Lane 5-6: 23 seconds；

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1603-14) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (ET1603-14) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse placenta tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse placenta tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-Phospho-EIF2S1 (S51) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT),Relative expression (RE)</span><br /><br />Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/200 dilution. <br /><br />A: Untreated human breast carcinoma tissue<br />B: λ-PPase treated human breast carcinoma tissue<br />C: Negative control<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT),Relative expression (RE)

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (ET1603-14) at 1/200 dilution.

    A: Untreated human breast carcinoma tissue
    B: λ-PPase treated human breast carcinoma tissue
    C: Negative control

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT),Relative expression (RE)</span><br /><br />Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/200 dilution. <br /><br />A: Untreated human colon carcinoma tissue<br />B: λ-PPase treated human colon carcinoma tissue<br />C: Negative control<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT),Relative expression (RE)

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (ET1603-14) at 1/200 dilution.

    A: Untreated human colon carcinoma tissue
    B: λ-PPase treated human colon carcinoma tissue
    C: Negative control

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of Phospho-EIF2S1 (S51) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

    Flow cytometric analysis of Phospho-EIF2S1 (S51) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1603-14, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1603-14" style="font-weight: bold;text-decoration: underline;">ET1603-14</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (ET1603-14) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-14) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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