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Western blot - eIF2 alpha antibody (ab26197)
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Western blot - eIF2 alpha antibody (ab26197)
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Immunocytochemistry/ Immunofluorescence - eIF2 alpha antibody (ab26197)
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Product Name
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eIF2 alpha antibody
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See all eIF2 alpha antibodies (10)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to eIF2 alpha
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human eIF2 alpha.
(Peptide available as ab26881.)
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Reacts with
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Hu, Ms, Dm
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Predicted to react
(species key)
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Rat
(Due to sequence homology)
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Tested applications
(see key)
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ICC/IF, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: 1/40. Perform 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).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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HeLa Whole Cell Lysate
Jurkat Whole Cell Lysate
A431 Whole Cell Lysate
NIH 3T3 Whole Cell Lysate
MEF1 Whole Cell Lysate
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Research areas
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Chromatin and Nuclear Signaling >> RNAi >> Eukaryotic Initiation factors (eIF's)
Chromatin and Nuclear Signaling >> DNA / RNA >> RNA Processing >> RNAi
Chromatin and Nuclear Signaling >> DNA / RNA >> Translation >> Regulation
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Relevance
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eIF2 alpha is a 36 kDa protein which is ubiquitously expressed in many cell types. The eIF2 protein, which is composed of three subunits (alpha, beta and gamma), is one of the key molecules in the initiation of translation. In mammalian cells, eIF2 alpha is phosphorylated at serine 51 (human EIF2 alpha, the equivalent residue in mouse is serine 52) by at least two kinases: the haem-controlled repressor (HCR) and the interferon inducible double stranded RNA-dependent protein kinase (PKR). Phosphorylation of eIF2 alpha blocks the GDP-GTP exchange activity of eIF2 beta, resulting in the suppression of protein synthesis. The phosphorylation of eIF2 alpha is an important regulatory process in protein synthesis.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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0.300 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this eIF2 alpha antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for eIF2 alpha antibody images, references, products related to ab26197 and other tools.
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eIF2 alpha antibody images:
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 Western blot - eIF2 alpha antibody (ab26197)
All lanes : eIF2 alpha 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.
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Western blot - eIF2 alpha antibody (ab26197)
All lanes : eIF2 alpha 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
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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).
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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
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References for eIF2 alpha antibody (ab26197)
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This product has been used in:
Farny NG et al. Metazoan stress granule assembly is mediated by P-eIF2alpha-dependent and -independent mechanisms. RNA 15:1814-21 (2009). WB; Fruit fly (Drosophila melanogaster). PubMed: 19661161
If you publish research using ab26197 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to eIF2 alpha
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eIF2 alpha antibody - more information
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