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Products:Epigenetics and Nuclear Signaling >> DNA / RNA >> Translation >> Regulation
Overview
Product name
Anti-EIF2S1 antibody
See all EIF2S1 products (14) ...
Description
Rabbit polyclonal to EIF2S1
Tested applications
ICC/IF, IHC-P, WBmore details
Cross reactivity
Reacts with
Mouse, Human, Fruit fly (Drosophila melanogaster)
Predicted to work with
Rat
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human eIF2 alpha.
(Peptide available as ab26881.)
Positive control
HeLa Whole Cell Lysate Jurkat Whole Cell Lysate A431 Whole Cell Lysate NIH 3T3 Whole Cell Lysate MEF1 Whole Cell Lysate
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Show applications key
Our Abpromise guarantee covers the use of ab26197 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use a concentration of 1 µg/ml
IHC-P: 1/40Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at an assay dependent concentration. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
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: EIF2AK3/PERK, GCN2, HRI and PKR. 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. In case of infection by vaccinia virus or rotavirus A, eIF2S1 phosphorylation state is modulated.
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. Co-localizes with NANOS3 in the stress granules.
Target information above from: UniProt accessionP05198
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- EIF 2 alpha antibody
- EIF 2 antibody
- EIF 2A antibody
see all
Database links
- Entrez Gene: 32617 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 1965 Human
- Entrez Gene: 13665 Mouse
- Entrez Gene: 54318 Rat
- Omim: 603907 Human
- SwissProt: P41374 Fruit fly (Drosophila melanogaster)
- SwissProt: P05198 Human
- SwissProt: Q6ZWX6 Mouse
see all
Anti-EIF2S1 antibody images:
Western blot - eIF2 alpha antibody (ab26197)
All lanes : Anti-EIF2S1 antibody (ab26197) at 1 µg/ml
Lane 1 : HeLa whole cell lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 36 kDa
Observed band size : 36 kDa
ab26197 detects a band of ~ 36 kDa in HeLa, Jurkat and A431 whole cell lysates. This corresponds to the predicted molecular weight for eIF2 alpha according to Swiss prot. A number of other bands are detected at ~ 48, 23 and 15 kDa. We do not know the identity of these bands and we predict they represent non-specific binding.
Western blot - eIF2 alpha antibody (ab26197)
All lanes : Anti-EIF2S1 antibody (ab26197) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 36 kDa
Observed band size : 36 kDa
Immunocytochemistry/ Immunofluorescence - eIF2 alpha antibody (ab26197)
ICC/IF image of ab26197 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab26197, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - eIF2 alpha antibody (ab26197)
Image courtesy of Human Protein Atlas
ab26197 staining eiF2 alpha in human small intestine. The paraffin embedded tissue was incubated with ab26197 (1/40 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab26197 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Further results for this antibody can be found at www.proteinatlas.org
References for Anti-EIF2S1 antibody (ab26197)
This product has been referenced in:
- Farny NGet al. Metazoan stress granule assembly is mediated by P-eIF2alpha-dependent and -independent mechanisms. RNA 15:1814-21 (2009). WB; Fruit fly (Drosophila melanogaster).Read more (PubMed: 19661161) »
See 1 publication for this product
Publishing research using ab26197? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"