Overview

  • Product name

    Anti-EIF2S1 antibody [EIF2a]
    See all EIF2S1 primary antibodies
  • Description

    Mouse monoclonal [EIF2a] to EIF2S1
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein (Human).

  • Epitope

    not mapped.
  • Positive control

    • Human Jurkat, CEM and HeLa cells, mouse 3T3L1 and rat PC-12 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab5369 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 Use a concentration of 5 µg/ml.
WB 1/500 - 1/1000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • 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).
  • Information by UniProt
  • Database links

  • 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

Images

  • All lanes : Anti-EIF2S1 antibody [EIF2a] (ab5369)

    Lane 1 : COS-7 whole cell extracts at 20 µg
    Lane 2 : MCF7 whole cell extracts at 20 µg
    Lane 3 : PC-12 at 20 µg
    Lane 4 : HeLa lysate
    Lane 5 : Jurkat
    Lane 6 : NIH/3T3
    Lane 7 : A-431

    Secondary
    Lanes 1-4 & 6-7 : Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution
    Lane 5 : Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate

    Predicted band size: 36 kDa

  • Immunohistochemical analysis of human colon carcinoma (right) compared to a negative control (left) using ab5369 at the dilution 1/10.

  • ICC/IF image of ab5369 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5369, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Ab5369 staining EIF2S1 in human placenta. Staining is localised to the cytoplasm.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

References

This product has been referenced in:

  • Cormerais Y  et al. Inhibition of the amino-acid transporter LAT1 demonstrates anti-neoplastic activity in medulloblastoma. J Cell Mol Med 23:2711-2718 (2019). Read more (PubMed: 30784173) »
  • Liu P  et al. Crizotinib-induced immunogenic cell death in non-small cell lung cancer. Nat Commun 10:1486 (2019). Read more (PubMed: 30940805) »
See all 36 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
60 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (gel 10%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 24 2012

Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (S2R+ cells)
Loading amount
25 µg
Specification
S2R+ cells
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycine)
Blocking step
normal horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Natalie Farny

Verified customer

Submitted Oct 22 2009

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cell line)
Loading amount
20 µg
Specification
HeLa cell line
Gel Running Conditions
Reduced Denaturing (12% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Mirjam Eckert

Verified customer

Submitted Apr 02 2009

Application
Western blot
Sample
Mouse Cell lysate - whole cell (RAW 264.7 murine macrophage)
Loading amount
10 µg
Specification
RAW 264.7 murine macrophage
Blocking step
Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Oct 23 2006

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