Phospho EIF2S1 (S51) antibody [E90] Recombinant (ab32157)

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

The antibody 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

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

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.
This product is a recombinant rabbit monoclonal antibody.

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
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

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 notes

Is unsuitable for 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 1/20000 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)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)

Immunohistochemical analysis of paraffin-embedded human liver carcinoma using ab32157 at 1/50 dilution.
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 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

ab32157 showing positive staining in Cervical carcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

ab32157 showing positive staining in Colonic adenocarcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 (phospho S51) antibody [E90] (ab32157)

ab32157 showing positive staining in Hepatocellular carcinoma tissue.

This product has been referenced in:

Collier AE et al. Translational control of a human CDKN1A mRNA splice variant regulates the fate of UVB-irradiated human keratinocytes. Mol Biol Cell 29:29-41 (2018). Read more (PubMed: 29118075) »
Meng C et al. PERK Pathway Activation Promotes Intracerebral Hemorrhage Induced Secondary Brain Injury by Inducing Neuronal Apoptosis Both in Vivo and in Vitro. Front Neurosci 12:111 (2018). Read more (PubMed: 29541018) »

See all 75 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 20 Abreviews or Q&A

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

Abcam guarantees this product to work in the species/application used in this Abreview.

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

Abcam guarantees this product to work in the species/application used in this Abreview.

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

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

Abcam user community

Verified customer

Submitted Dec 13 2016

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

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

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

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

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

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 31 2014

1-10 of 20 Abreviews or Q&A

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

Overview

Properties

Associated products

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Target

Images

Datasheets and documents

References

This product has been referenced in:

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]

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]

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]

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