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EIF2S1 Recombinant Rabbit Monoclonal Antibody [PSH04-29]

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Specification

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Catalog# HA722112

EIF2S1 Recombinant Rabbit Monoclonal Antibody [PSH04-29]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

EIF2S1 Recombinant Rabbit Monoclonal Antibody [PSH04-29]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human EIF2S1 aa 1-315 / 315.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

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

Molecular Weight

Predicted band size: 36 kDa

Positive Control

MCF7 cell lysate, HepG2 cell lysate, HeLa cell lysate, COS-1 cell lysate, A549 cell lysate, RAW264.7 cell lysate, C6 cell lysate, mouse kidney tissue lysate, mouse spleen tissue lysate, rat kidney tissue lysate, rat spleen tissue lysate, human breast tissue, mouse breast tissue, rat breast tissue, HepG2.

Conjugation

unconjugated

Clone Number

PSH04-29

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Eukaryotic translation initiation factor 2 subunit 1 (eIF2α) is a protein that in humans is encoded by the EIF2S1 gene. The protein encoded by this gene is the alpha (α) subunit of the translation initiation factor eIF2 protein complex which catalyzes an early regulated step of protein synthesis initiation, promoting the binding of the initiator tRNA (Met-tRNAiMet) to 40S ribosomal subunits. Binding occurs as a ternary complex of methionyl-tRNA, eIF2, and GTP. eIF2 is composed of 3 nonidentical subunits, alpha (α, 36 kD, this article), beta (β, 38 kD), and gamma (γ, 52 kD). The rate of formation of the ternary complex is modulated by the phosphorylation state of eIF2α. Phosphorylation of eIF2α by EIF-2 kinases plays a key role in regulating the integrated stress response.

Background References

1. Dang TT et al. Phosphorylation of EIF2S1 (eukaryotic translation initiation factor 2 subunit alpha) is indispensable for nuclear translocation of TFEB and TFE3 during ER stress. Autophagy. 2023 Jul

2. Shi JX et al. MiR-3074-5p Regulates Trophoblasts Function via EIF2S1/GDF15 Pathway in Recurrent Miscarriage. Reprod Sci. 2023 Dec

Subcellular Location

Cytoplasm, Stress granule, cytosol, Mitochondrion.

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

  • Western blot analysis of EIF2S1 on different lysates with Rabbit anti-EIF2S1 antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: COS-1 cell lysate<br />Lane 5: A549 cell lysate<br />Lane 6: RAW264.7 cell lysate<br />Lane 7: C6 cell lysate<br />Lane 8: Mouse kidney tissue lysate<br />Lane 9: Mouse spleen tissue lysate<br />Lane 10: Rat kidney tissue lysate<br />Lane 11: Rat spleen tissue lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 36 kDa<br />Observed band size: 36 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<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/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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.

    Western blot analysis of EIF2S1 on different lysates with Rabbit anti-EIF2S1 antibody (HA722112) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

    Lane 1: MCF7 cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: COS-1 cell lysate
    Lane 5: A549 cell lysate
    Lane 6: RAW264.7 cell lysate
    Lane 7: C6 cell lysate
    Lane 8: Mouse kidney tissue lysate
    Lane 9: Mouse spleen tissue lysate
    Lane 10: Rat kidney tissue lysate
    Lane 11: Rat spleen tissue lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 59 seconds; ECL: K1801;
    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 (HA722112) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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 breast tissue with Rabbit anti-EIF2S1 antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</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/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</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 human breast tissue with Rabbit anti-EIF2S1 antibody (HA722112) 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 (HA722112) 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 breast tissue with Rabbit anti-EIF2S1 antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</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/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</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 breast tissue with Rabbit anti-EIF2S1 antibody (HA722112) 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 (HA722112) 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 rat breast tissue with Rabbit anti-EIF2S1 antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</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/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</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 rat breast tissue with Rabbit anti-EIF2S1 antibody (HA722112) 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 (HA722112) 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.

  • Immunocytochemistry analysis of HepG2 cells labeling EIF2S1 with Rabbit anti-EIF2S1 antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-EIF2S1 antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling EIF2S1 with Rabbit anti-EIF2S1 antibody (HA722112) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-EIF2S1 antibody (HA722112) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HepG2 cells labeling EIF2S1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling EIF2S1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722112, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • EIF2S1 was immunoprecipitated from 0.2 mg HepG2 cell lysate with <a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HepG2 cell lysate (input)<br />Lane 2: <a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a> IP in HepG2 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722112" style="font-weight: bold;text-decoration: underline;">HA722112</a> in HepG2 cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 2 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    EIF2S1 was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA722112 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722112 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HepG2 cell lysate (input)
    Lane 2: HA722112 IP in HepG2 cell lysate
    Lane 3: Rabbit IgG instead of HA722112 in HepG2 cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 2 seconds; ECL: K1801

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Citation