Name: Anti-eIF2A antibody [EPR11042]
Brand: RabMAb
SKU: ab169528
Rating: 5 (2 reviews)

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Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR11042] to eIF2A
Suitable for: WB, IHC-P, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-eIF2A antibody [EPR11042]
See all eIF2A primary antibodies
Description

Rabbit monoclonal [EPR11042] to eIF2A
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt or IP
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide corresponding to Human eIF2A aa 450-550.
Positive control
- WB: Molt4, Ramos, HAP1, PC-12, and NIH/3T3 lysates. IHC-P: Human pancreas, Human prostatic hyperplasia, Human lung cancer, Mouse cerebrum, and Rat cerebrum tissues. ICC/IF: HeLa cells, MCF7 cells.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

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

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR11042
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- Ramos whole cell lysate (ab3955)
- MOLT4 whole cell lysate (ab7912)
Recombinant Protein
- Recombinant Human eIF2A protein (ab95932)

Our Abpromise guarantee covers the use of ab169528 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

1/1000 - 1/5000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).

IHC-P

1/50 - 1/100.

Perform heat mediated antigen retrieval.

ICC/IF

1/100.

Application notes

Is unsuitable for Flow Cyt or IP.

Function

Functions in the early steps of protein synthesis of a small number of specific mRNAs. Acts by directing the binding of methionyl-tRNAi to 40S ribosomal subunits. In contrast to the eIF-2 complex, it binds methionyl-tRNAi to 40 S subunits in a codon-dependent manner, whereas the eIF-2 complex binds methionyl-tRNAi to 40 S subunits in a GTP-dependent manner. May act by impiging the expression of specific proteins.
Tissue specificity

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

Belongs to the WD repeat EIF2A family.
Contains 3 WD repeats.
Target information above from: UniProt accession Q9BY44 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 83939 Human
- Entrez Gene: 229317 Mouse
- Entrez Gene: 502531 Rat
- Omim: 609234 Human
- SwissProt: Q9BY44 Human
- SwissProt: Q8BJW6 Mouse
- SwissProt: P68101 Rat
- Unigene: 655782 Human
- Unigene: 21617 Mouse
- Unigene: 198593 Rat
see all
Alternative names
- 65 kDa eukaryotic translation initiation factor 2A antibody
- CDA 02 antibody
- CDA02 antibody
- EIF 2 antibody
- EIF 2A antibody
- eIF-2A antibody
- EIF2 antibody
- eif2a antibody
- EIF2A_HUMAN antibody
- Eukaryotic translation initiation factor 2A 65kDa antibody
- Eukaryotic translation initiation factor 2A antibody
- MST089 antibody
- MSTP004 antibody
- MSTP089 antibody
see all

Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: eIF2A knockout HAP1 cell lysate (20 µg)
Lane 3: Ramos cell lysate (20 µg)
Lane 4: MOLT4 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab169528 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab169528 was shown to specifically react with eIF2A when eIF2A knockout samples were used. Wild-type and eIF2A knockout samples were subjected to SDS-PAGE. ab169528 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling eIF2A with Purified ab169528 at 1:100 dilution (4.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)

Anti-eIF2A antibody [EPR11042] (ab169528) at 1/1000 dilution (Purified) + NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 65 kDa
Observed band size: 65 kDa
Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)

All lanes : Anti-eIF2A antibody [EPR11042] (ab169528) at 1/1000 dilution (Purified)

Lane 1 : Ramos (Human Burkitt's lymphoma Blymphocyte) whole cell lysates
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) 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: 65 kDa
Observed band size: 65 kDa
Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)

ab169528 staining eIF2A in wild-type HAP1 cells (top panel) and EIF2A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab169528 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling eIF2A with purified ab169528 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunohistochemical analysis of paraffin embedded Human pancreas tissue labeling eIF2A with unpurified ab169528 antibody at 1/50.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)

Immunohistochemical analysis of paraffin embedded Human prostatic hyperplasia tissue labeling eIF2A with unpurified ab169528 antibody at 1/50.
Anti-eIF2A antibody [EPR11042] (ab169528)

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheet

Publishing research using ab169528? Please let us know so that we can cite the reference in this datasheet.

ab169528 has been referenced in 7 publications.

Zhang J et al. Inhibition of the SIRT1 signaling pathway exacerbates endoplasmic reticulum stress induced by renal ischemia/reperfusion injury in type 1 diabetic rats. Mol Med Rep 21:695-704 (2020). PubMed: 31974604
Lin Y et al. Exosomes derived from HeLa cells break down vascular integrity by triggering endoplasmic reticulum stress in endothelial cells. J Extracell Vesicles 9:1722385 (2020). PubMed: 32128072
Li D et al. Lobaplatin promotes 125I-induced apoptosis and inhibition of proliferation in hepatocellular carcinoma by upregulating PERK-eIF2a-ATF4-CHOP pathway. Cell Death Dis 10:744 (2019). PubMed: 31582720
Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
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). PubMed: 29541018
Zhang Z et al. METTL3-mediated N6-methyladenosine mRNA modification enhances long-term memory consolidation. Cell Res N/A:N/A (2018). WB ; Mouse . PubMed: 30297870
Golovko A et al. The eIF2A knockout mouse. Cell Cycle 15:3115-3120 (2016). PubMed: 27686860

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Application

Western blot

Sample

Human Cell lysate - whole cell (Dopaminergic Neurons)

Gel Running Conditions

Reduced Denaturing (NuPage 4-12% MES buffer)

Loading amount

30 µg

Specification

Dopaminergic Neurons

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 Oct 15 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (MDA231 cells)

Gel Running Conditions

Reduced Denaturing (10% Tris-Glycine Gel)

Loading amount

50000 cells

Specification

MDA231 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Aug 16 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Recombinant Anti-eIF2A antibody [EPR11042] (ab169528)

Key features and details

Overview

Product name

Description

Host species

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

Isotype control

Positive Controls

Recombinant Protein

Applications

Target

Function

Tissue specificity

Sequence similarities

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (7)

Customer reviews and Q&As

Western blot abreview for Anti-eIF2A antibody [EPR11042]

Western blot abreview for Anti-eIF2A antibody [EPR11042]