Anti-EIF2S1 antibody [EIF2a] (ab5369)

Reviews (4)Q&A (6)References (31)

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
    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

Images

  • Western blot - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    Western blot - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 antibody [EIF2a] (ab5369)

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

  • Immunocytochemistry/ Immunofluorescence - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    Immunocytochemistry/ Immunofluorescence - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EIF2S1 antibody [EIF2a] (ab5369)
    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. The glutamine transporter ASCT2 (SLC1A5) promotes tumor growth independently of the amino acid transporter LAT1 (SLC7A5). J Biol Chem 293:2877-2887 (2018). Read more (PubMed: 29326164) »
  • Xu Y  et al. Chrysin induces cell growth arrest, apoptosis, and ER stress and inhibits the activation of STAT3 through the generation of ROS in bladder cancer cells. Oncol Lett 15:9117-9125 (2018). Read more (PubMed: 29805643) »
See all 31 Publications for this product

Publishing research using ab5369? 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 10 Abreviews or Q&A

Western blot abreview for Anti-EIF2S1 antibody [EIF2a]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
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
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Abcam user community

Verified customer

Submitted Jan 24 2012

Question

Dear, My name is Tânia Lourenço and i am a master's student in Molecular and Cell Biology. I am contacting you because i have some questions about Fyn (phospho Y530) antibody (ab5369). I would like to know if the antibody can recognize other src proteins and if you could please tell me for sure that the antibody recognizes only the Fyn and no other proteins.  I thank you for your time. Yours sincerely,

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Answer

Thank you for your enquiry. I am sorry it has taken some time to confirm this information with the originator for this antibody. I am sorry to confirm that the only cross reactivity data we have for ab5369 Anti-EIF2S1 antibody [EIF2a] is that it recognizes eIF2-alpha from human, mouse, and rat samples. The epitope information is proprietary for this particular product and I am sorry we are not able to provide the details on this occasion. I would like to reassure you that this antibody is covered by our Abpromise guarantee in the tested applications and species listed on the datasheet. This means we are pleased to provide a refund, credit note or free of charge replacement if the antibody is not working in the tested applications and species. The guarantee period is for 6 months after purchase. There are 3 references available on the use of this antibody that I hope may be helpful to you: Ramos-Casteneda et al. (2005) J. Biol. Chem. 280 (10) 9467-9473 Trottier, M. et al. (2005) Virology 333(2): 215-225 Zhang, P. et al. (2002) Molec. Cell. Biol. 22 (11): 3864-3874 I am sorry we have no more data for your requriements on this occasion. Please do not hesitate to contact me if you have any other questions.

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Western blot abreview for Anti-EIF2S1 antibody [EIF2a]

 Inconclusive
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
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
Read More

Dr. Natalie Farny

Verified customer

Submitted Oct 22 2009

Western blot abreview for Anti-EIF2S1 antibody [EIF2a]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
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
Read More

Dr. Mirjam Eckert

Verified customer

Submitted Apr 02 2009

Question

I have purchased a number of antibodies from Abcam and they have all been good. I have couple of questions which i would like you to assist me with, as i have checked your website and found no answers. The Rabbit polyclonal eIF4E antibody (ab1126) does it detect total eIF4E ie the non phospho and the phospho or just the non phospho form The Mouse Monoclonal eIF2a antibody (ab5369) does it detect total eIF2a ie the non phospho and the phospo or just the non phospho form

Read More
Answer

Thank you for your enquiry. I can confirm that both these eIF2a antibodies (ab1126, ab5369)will detect both phosphorylated and de-phosphorylated forms of the protein. I hope this information is helpful. Should you have any further questions, please do not hesitate to contact us again.

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Western blot abreview for Anti-EIF2S1 antibody [EIF2a]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
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%
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Abcam user community

Verified customer

Submitted Oct 23 2006

Question

No, Swissprot, EBI's and NCBI's websites are riddled with mistakes and faulty information. So, here's the link you should look at: http://www.chem.qmul.ac.uk/iubmb/misc/trans.html It is IUBMB that has the naming rights to translation factors, no one else. Thank you for your time and letting help you correct other's mistakes.

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Answer

Thank you for this information and clarification. I have changed the online datasheet to reflect the fact that the antibody is raised against eIF2 alpha. Thank you again for your assistance.

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Question

The antibody products stated to be anti eIF-2A really against eIF2alpha. They are two entirely different proteins. ab4837 ab5369 ab19288

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Answer

Thank you for your enquiry. Our designation comes directly from Swiss-Prot where the synonyms for the protein are as follows: Eukaryotic translation initiation factor 2 alpha subunit eIF-2-alpha EIF-2alpha EIF-2A I believe the datasheet is correct in identifying the protein that the antibody is raised against. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

I see that you have three antibodies to eIF2A. I'm particularly interested in an antibody that reacts with the c terminal region of eIF2A (near aa 214-234). antibodies ab4837 and ab19288 apparently react with the n-terminal region (viz., phosphorylated S52 or s51(?)). antibody ab5369, on trhe other hand, was developed against the whole protein, but do you have any ideas as to the location of the epitope. If the epitope of the latter antibody is located in the c-terminal half to eIF2A, it may suit my needs. please let me know if you have any ideas as to location of the epitope of ab5369. thank you

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Answer

We have received the following information from the source of this antibody: "The Mouse (monoclonal) anti-eIF2alpha antibody ab5369 clone EIF2alpha was made using recombinant protein. There is unfortunately no information on the epitope or region the antibody recognizes. The following references cite the antibody: Ramos-Castaneda J, et al (2005) J. Biol. Chem. 280(10):9467-9473. Trottier MD, et al (2005) Virology 333:215-225. Zhang, et al (2002) Mol. Cell. Biol. 22(11):3864-3874. Please do not hesitate to contact us again if you require further information,

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Question

I am interested in the mouse monoclonal Ig against eIF2alpha (ab5369). A western blot prepared from cell lysates is shown on the data sheet (HeLa) and while the Ig concentration used for the western blot is shown, the amount of loaded target is not discussed. Was the total protein loaded for each lysate determined before loading the gel? What is the sensitivity of the Ig? How much lysate was loaded (total protein - ug's/ well; or in terms of starting cell #s). We are interested in the antibody and would use it in assays where we have 5 - 15 micrograms of TOTAL HeLa cell lysate loaded/ well (lysate prepared from 5 x10EE4 - 1 x 10EE5 cells). We don't want to spend money on an Ig that is insensitive and cannot detect eIF2alpha in these assays.

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Answer

I enquired with the supplier of ab5369 and they were able to provide me with the following information: approximately 30-40 micrograms of lysate was loaded. If you have any more questions, do not hesitate to contact us again.

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