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    products/primary-antibodies/eif2a-antibody-epr11042-ab169528.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-eIF2A antibody [EPR11042] (ab169528)

  • Datasheet
  • SDS
Reviews (5) Submit a question References (14)

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Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (ab169528)
  • 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

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Overview

  • 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.

Properties

  • 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

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • Translation
    • Regulation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • RNAi
    • Epigenetics and Nuclear Signaling
    • RNAi
    • Eukaryotic Initiation factors (eIF's)

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-eIF2A antibody [EPR11042] (ab214876)
    • Alexa Fluor® 647 Anti-eIF2A antibody [EPR11042] (ab215231)
    • HRP Anti-eIF2A antibody [EPR11042] (ab215342)
    • Anti-eIF2A antibody [EPR11042] - BSA and Azide free (ab236012)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Ramos whole cell lysate (ab3955)
    • MOLT-4 whole cell lysate (ab7912)
  • Recombinant Protein

    • Recombinant Human eIF2A protein (ab95932)

Applications

The Abpromise guarantee

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 (3)
1/1000 - 1/5000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
IHC-P (1)
1/50 - 1/100.

Perform heat mediated antigen retrieval. 

ICC/IF
1/100.
Notes
WB
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.

Target

  • 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
    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

Images

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

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (14)

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

ab169528 has been referenced in 14 publications.

  • Liu Z  et al. Overexpression of thioredoxin reductase 1 can reduce DNA damage, mitochondrial autophagy and endoplasmic reticulum stress in Parkinson's disease. Exp Brain Res 239:475-490 (2021). PubMed: 33230666
  • Meng X  et al. Endoplasmic reticulum stress promotes epithelial-mesenchymal transition via the PERK signaling pathway in paraquat-induced pulmonary fibrosis. Mol Med Rep 24:N/A (2021). PubMed: 34036384
  • Martin A  et al. Gut microbiota mediate the FGF21 adaptive stress response to chronic dietary protein-restriction in mice. Nat Commun 12:3838 (2021). PubMed: 34158480
  • Bao J  et al. Pharmacological Disruption of Phosphorylated Eukaryotic Initiation Factor-2a/Activating Transcription Factor 4/Indian Hedgehog Protects Intervertebral Disc Degeneration via Reducing the Reactive Oxygen Species and Apoptosis of Nucleus Pulposus Cells. Front Cell Dev Biol 9:675486 (2021). PubMed: 34164397
  • Rong K  et al. Alendronate Alleviated Femoral Head Necrosis and Upregulated BMP2/EIF2AK3/EIF2A/ATF4 Pathway in Liquid Nitrogen Treated Rats. Drug Des Devel Ther 15:1717-1724 (2021). PubMed: 33935494
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Immunocytochemistry abreview for Anti-eIF2A antibody [EPR11042]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cultured Cells (U937 cells)
Permeabilization
Yes - 0.1% Triton-X100 in PBS 10'
Specification
U937 cells
Blocking step
Bloxall as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 07 2022

Western blot abreview for Anti-eIF2A antibody [EPR11042]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (CT26 cells)
Gel Running Conditions
Reduced Denaturing (4-12% Tris-Bis gel)
Loading amount
40 µg
Specification
CT26 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 10 2022

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-eIF2A antibody [EPR11042]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Panc1 Tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA ph9
Permeabilization
No
Specification
Panc1 Tumor
Blocking step
Bloxall as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 27 2021

Western blot abreview for Anti-eIF2A antibody [EPR11042]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 15 2020

Western blot abreview for Anti-eIF2A antibody [EPR11042]

Excellent
Abreviews
Abreviews
abreview image
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%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 16 2016

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