Anti-EIF2S1 antibody [EIF2a] (ab5369)
- Datasheet
- References (31)
- Protocols
Overview
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Product nameAnti-EIF2S1 antibody [EIF2a]
See all EIF2S1 primary antibodies -
DescriptionMouse monoclonal [EIF2a] to EIF2S1
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Host speciesMouse
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Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Recombinant full length protein (Human).
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Epitopenot mapped.
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Positive control
- Human Jurkat, CEM and HeLa cells, mouse 3T3L1 and rat PC-12 cells.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferPreservative: 0.1% Sodium Azide
Constituents: PBS, pH 7.4 -
Concentration information loading...
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PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEIF2a
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IsotypeIgG1
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Research areas
Associated products
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Alternative Products
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Positive Controls
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 |
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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
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FunctionFunctions 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.
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Sequence similaritiesBelongs to the eIF-2-alpha family.
Contains 1 S1 motif domain. -
Post-translational
modificationsSubstrate 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 localizationCytoplasmic 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
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Database links
- Entrez Gene: 1965 Human
- Entrez Gene: 13665 Mouse
- Entrez Gene: 54318 Rat
- Omim: 603907 Human
- SwissProt: P05198 Human
- SwissProt: Q6ZWX6 Mouse
- SwissProt: P68101 Rat
- Unigene: 151777 Human
see all -
Alternative names
- EIF 2 alpha antibody
- EIF 2 antibody
- EIF 2A antibody
see all
Images
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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)
Immunohistochemical analysis of human colon carcinoma (right) compared to a negative control (left) using ab5369 at the dilution 1/10.
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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.
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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.
Protocols
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) »