EIF2S1 ELISA Kit (pS51) is a single-wash 90-min Simplestep used to quantify EIF2S1 (pS51) with a sensitivity of 9 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
EIF2S1 ELISA Kit (pS51) is a single-wash 90-min Simplestep used to quantify EIF2S1 (pS51) with a sensitivity of 9 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Sample | n | C.V. |
---|---|---|
Sample Extract | n 8 | C.V. 3.8 |
Sample | n | C.V. |
---|---|---|
Sample Extract | n 3 | C.V. 4.7 |
Sample type | Average % | Range |
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Sample type Cell Lysate | Average % = 106 | Range 99 - 114 % |
EIF2S1 ELISA kit (ab282296) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of EIF2S1 protein in human cell extract samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate EIF2S1 with 9 units/mL sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
EIF2S1, Eukaryotic translation initiation factor 2 subunit 1, plays a key role in the initiation of protein synthesis. Dysregulation of EIF2S1 plays a part in the development of type II diabetes and ischemic injury. The immunogen from which the antibodies in this kit were raised represents the full-length mature protein. This protein shares homology across Mammalia, and therefore is expected to cross-react with many species. The protein standard in this product is recombinant human EIF2S1 ligated with peptide containing phosphorylated serine 51.
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EIF2S1 also called eIF2α is a subunit of the eukaryotic translation initiation factor 2 (eIF2) complex. This protein plays an important role in the initiation of protein synthesis by regulating the binding of the initiator methionyl-tRNA to the 40S ribosomal subunit. Its molecular weight is approximately 36 kDa. EIF2S1 is widely expressed in various tissues providing a fundamental function in protein biosynthesis across different cell types.
EIF2α acts as a regulatory component of the eIF2 complex which is central to initiating translation in eukaryotic cells. Phosphorylation of eIF2α leads to inhibition of the exchange of GDP to GTP on eIF2 effectively reducing global protein synthesis while allowing the translation of specific mRNAs involved in stress responses. The eIF2α forms part of the eIF2 complex working closely with other subunits to exert its regulatory functions.
EIF2α is involved in the integrated stress response (ISR) and other protein synthesis pathways. The phosphorylation state of eIF2α is important in ISR linking it to proteins such as PERK and GCN2 which respond to various stress signals. Phospho-eIF2α plays a significant role in modulating the translation response to cellular stress ensuring cells can adapt or initiate apoptosis depending on the conditions.
EIF2α connects to diseases like Alzheimer's and diabetes. Alterations in phosphorylation of eIF2α are linked to endoplasmic reticulum stress in Alzheimer's affecting protein homeostasis. Connecting to diabetes dysregulated eIF2α signaling interferes with insulin synthesis and secretion. Proteins like ATF4 also intersect with eIF2α in these diseases highlighting eIF2α's involvement in critical pathological processes.
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Cells were treated with 50 nM pervanadate for 30 minutes, scraped and buffer exchanged into PBS + 20 mM NaF, followed by protein extraction. Mock control cells accompanied cell treatment. Concentrations of EIF2S1(pS51) were measured in duplicate and interpolated against the standard curve. Results show the mean EIF2S1(pS51) concentration was 68.55 pg/mL in 7.5 ug/mL HeLa control cells, 876.09 pg/mL in 7.5 ug/mL HeLa + Calyculin, 173.27 pg/mL in 12.5 ug/mL NIH-3T3 control, 919.27 pg/mL in 12.5 ug/mL NIH-3T3 + Calyculin, 112.07 pg/mL in 12.5 ug/mL C6 control, and 813.39 pg/mL in 7.5 ug/mL C6 + Calyculin.
The concentrations of EIF2S1(pS51) were measured in duplicate and interpolated from the EIF2S1(pS51) standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean EIF2S1(pS51) concentration was determined to be 3,070.84 pg/mL in HeLa + Calyculin cell extract, 2,802.47 pg/mL in NIH-3T3 + Calyculin cell extract and 2,650.15 pg/mL in C6 + Calyculin cell extract.
The EIF2S1(pS51) standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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