Phospho-EIF2S1 (S52) and Total EIF2S1 ELISA Kit (ab279775)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
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Product name
Phospho-EIF2S1 (S52) and Total EIF2S1 ELISA Kit
See all EIF2S1 kits -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
Phospho-EIF2S1 (S52) and Total EIF2S1 ELISA Kit (ab279775) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated EIF2S1 (S52) protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-EIF2S1 and total EIF2S1. An anti-pan EIF2S1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and EIF2S1 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, biotinylated anti-phospho-eIF-2A (S52) antibody is added to detect phosphorylated EIF2S1. In the remaining wells, rabbit anti-pan-eIF-2A antibody is used to detect pan EIF2S1. After washing away unbound antibody, HRP-Streptavidin or HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of EIF2S1 (S52) or pan EIF2S1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 20X Wash Buffer Concentrate 1 x 25ml 2X Cell lysate buffer 1 x 10ml 300X Streptavidin-Conjugated HRP 1 vial 5X Assay Diluent 1 x 15ml Biotinylated anti-phospho-EIF2S1 (S52)-antibody 1 vial HRP-conjugated anti-rabbit IgG concentrate (1000X) 1 vial Pan-EIF2S1 Coated Microplate 1 unit Positive Control - HeLa cell lysate 1 vial Rabbit anti-pan-EIF2S1 detection antibody 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
Research areas
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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
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 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
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Alternative names
- EIF 2
- EIF 2 alpha
- EIF 2A
see all -
Database links
- Entrez Gene: 1965 Human
- Omim: 603907 Human
- SwissProt: P05198 Human
- Unigene: 151777 Human
Images
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Positive Control
HeLa cells were treated with Calyculin A and TNF-a.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
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Jurkat cells were treated with/without Calyculin A and Pervanadate.Jurkat cells were treated or untreated with Calyculin A and Pervanadate.
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Jurkat cells were treated with/without Calyculin A and Pervanadate.Jurkat cells were treated or untreated with Calyculin A and Pervanadate.
