Name: Anti-EIF2S1 (phospho S51) antibody [E90]
Brand: RabMAb
SKU: ab32157
Rating: 4 (24 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E90] to EIF2S1 (phospho S51)
Suitable for: WB, IHC-P, Dot blot
Reacts with: Rat, Human, Neurospora crassa

Carrier Free

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: WB, IHC-P, Dot blotmore details
Unsuitable for: Flow Cyt,ICC or IP
Species reactivity

Reacts with: Rat, Human, Neurospora crassa
Predicted to work with: Mouse, Cow, Pig, Drosophila melanogaster, Monkey, Plants, African green monkey
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- IHC-P: Human liver carcinoma, breast carcinoma, cervical carcinoma, colon adenocarcinoma and hepatocellular carcinoma tissues. WB: HeLa cells treated with Clyculin A and phosphatase whole cell lysate; PC-12 cell lysate. 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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E90
Isotype

IgG
Research areas

Alternative Versions
- Anti-EIF2S1 (phospho S51) antibody [E90] - BSA and Azide free (ab214434)
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

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
WB	(19)	1/1000 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P	(1)	1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Dot blot		1/500.

Notes
WB 1/1000 - 1/10000. 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.
Dot blot 1/500.

Application notes

Is unsuitable for Flow Cyt,ICC or IP.

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).
Target information above from: UniProt accession P05198 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 327694 Cow
- Entrez Gene: 32617 Drosophila melanogaster
- Entrez Gene: 1965 Human
- Entrez Gene: 13665 Mouse
- Entrez Gene: 54318 Rat
- Omim: 603907 Human
- SwissProt: P41374 Drosophila melanogaster
- SwissProt: P05198 Human
- SwissProt: Q6ZWX6 Mouse
- SwissProt: P68101 Rat
- Unigene: 3157 Drosophila melanogaster
- Unigene: 151777 Human
- Unigene: 196220 Mouse
- Unigene: 1488 Rat
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

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

All lanes : Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157) at 1/1000 dilution

Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) Whole cell lysates
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 5 ug/ml tunicamycin for 18 hours whole cell lysates
Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 5 ug/ml tunicamycin for 18 hours whole cell lysates. Then the membrane was incubated with alkaline phosphatase.
Lane 4 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 5 ug/ml tunicamycin for 18 hours whole cell lysates. Then the membrane was incubated with lambda phosphatase.

Lysates/proteins at 15 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

Exposure time: 180 seconds

Blocking/Diluting buffer and concentration 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

Immunohistochemical analysis of paraffin-embedded human lung cancer sections labeling EIF2S1 with ab32157 at 1/4000 dilution (0.37 μg/mL). Hematoxylin was used as counterstain. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Antigen retrieval was heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Positive staining on human lung cancer without alkaline phosphatase treatment (image A). No staining on human lung cancer with alkaline phosphatase treatment (image B)
The section was incubated with ab32157 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Dot Blot - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

Dot blot analysis of EIF2S1 (pS51) phospho peptide (Lane 1), EIF2S1 non-phospho peptide (Lane 2) with ab32157 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

Immunohistochemical analysis of paraffin-embedded mouse pancreatic cancer sections labeling EIF2S1 with ab32157 at 1/4000 dilution (0.37 μg/mL). Hematoxylin was used as counterstain. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Antigen retrieval was heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Positive staining on mouse pancreatic cancer without alkaline phosphatase treatment (image A). No staining on mouse pancreatic cancer with alkaline phosphatase treatment (image B)
The section was incubated with ab32157 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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.
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)

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)

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)

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)

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)

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

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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

ab32157 has been referenced in 179 publications.

Popovic R et al. Combined Transcriptomic and Proteomic Analysis of Perk Toxicity Pathways. Int J Mol Sci 22:N/A (2021). PubMed: 33925631
Huang Y et al. Elucidation of the mechanism of NEFA-induced PERK-eIF2a signaling pathway regulation of lipid metabolism in bovine hepatocytes. J Steroid Biochem Mol Biol 211:105893 (2021). PubMed: 33819629
Arioka Y et al. Chromosome 22q11.2 deletion causes PERK-dependent vulnerability in dopaminergic neurons. EBioMedicine 63:103138 (2021). PubMed: 33341442
Tian AL et al. Lysosomotropic agents including azithromycin, chloroquine and hydroxychloroquine activate the integrated stress response. Cell Death Dis 12:6 (2021). PubMed: 33414432
Zyryanova AF et al. ISRIB Blunts the Integrated Stress Response by Allosterically Antagonising the Inhibitory Effect of Phosphorylated eIF2 on eIF2B. Mol Cell 81:88-103.e6 (2021). PubMed: 33220178

View all Publications for this product

Customer reviews and Q&As

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1-10 of 26 Abreviews or Q&A

Application

Western blot

Sample

Mouse Tissue lysate - whole (Heart, Liver)

Gel Running Conditions

Reduced Denaturing (12% TGX)

Loading amount

20 µg

Specification

Heart, Liver

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 22 2021

Application

Western blot

Sample

Xenopus laevis Cell lysate - whole cell (Unfertilised Xenopus oocytes, stage V-VI)

Gel Running Conditions

Reduced Denaturing (4-12% Bis-Tris)

Loading amount

0.5 cells

Specification

Unfertilised Xenopus oocytes, stage V-VI

Blocking step

BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Rowena Martin

Verified customer

Submitted Nov 17 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Human Primary Fibroblasts)

Permeabilization

Yes - 0.1% Triton 1h

Specification

Human Primary Fibroblasts

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 21 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (Human Primary Fibroblasts)

Gel Running Conditions

Reduced Denaturing (12% TGX)

Loading amount

10 µg

Treatment

different treatments

Specification

Human Primary Fibroblasts

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Oct 21 2020

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

Mr. Kyung noh Yoon

Verified customer

Submitted May 09 2019

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

Mr. Jin Ik Kim

Verified customer

Submitted Apr 20 2019

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

Abcam user community

Verified customer

Submitted Jul 07 2018

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

Abcam user community

Verified customer

Submitted Jul 25 2017

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

Miss. Berta Anuncibay

Verified customer

Submitted May 04 2017

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

Abcam user community

Verified customer

Submitted Feb 14 2017

1-10 of 26 Abreviews or Q&A

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For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (179)

Customer reviews and Q&As

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

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

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

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

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

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

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

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

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

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

Post-translational
modifications