Name: Anti-eIF2A antibody [EPR11042]
Brand: RabMAb
SKU: ab169528
Rating: 5 (1 reviews)

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, and BxPC-3 cell lysates; HeLa cells, PC-12 whole cell lysates, NIH/3T3 whole cell lysates; IHC-P: Human pancreas, Human prostatic hyperplasia, Human lung cancer, Mouse cerebrum, and Rat cerebrum tissues. ICC/IF: HeLa cells, HAP1 wildtype and HAP1-EIF2A knockout cells.
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
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 long term. Avoid freeze / thaw cycle.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR11042
Isotype

IgG
Research areas

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
WB		1/1000 - 1/5000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
IHC-P		1/50 - 1/100. See IHC antigen retrieval protocols.
ICC/IF		Use a concentration of 1 µg/ml.

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)

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

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

Lane 1 : Molt4 cell lysate
Lane 2 : Ramos cell lysate
Lane 3 : BxPC-3 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 65 kDa
Observed band size: 65 kDa
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.

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

Datasheet

This product has been referenced in:

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) »
Zhang Z et al. METTL3-mediated N6-methyladenosine mRNA modification enhances long-term memory consolidation. Cell Res N/A:N/A (2018). WB ; Mouse . Read more (PubMed: 30297870) »

See all 3 Publications for this product

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

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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%

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Verified customer

Submitted Aug 16 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-eIF2A antibody [EPR11042] (ab169528)

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This product has been referenced in:

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Western blot abreview for Anti-eIF2A antibody [EPR11042]