ELISA/assay

Phospho-eIF2alpha (Ser51) Colorimetric Cell-Based ELISA Kit

MBS9501438

2 x96 Wells

535 USD

ELISA Kit

Phospho-eIF2alpha (Ser51) Colorimetric Cell-Based ELISA Kit

[eIF2alpha]

[eIF-2-alpha; EIF-2A; EIF-2alpha; EIF2S1; Eukaryotic translation initiation factor 2 alpha subunit; Eukaryotic translation initiation factor 2 subunit 1; IF2A]

[eukaryotic translation initiation factor 2 subunit 1; Eukaryotic translation initiation factor 2 subunit 1; eukaryotic translation initiation factor 2 subunit 1; eukaryotic translation initiation factor 2 subunit alpha; Eukaryotic translation initiation factor 2 subunit alpha; eIF-2-alpha; eIF-2A; eIF-2alpha]

[eIF2alpha]

[EIF2S1]

[EIF2S1; EIF2S1; EIF2; EIF-2; EIF2A; EIF-2A; EIF-2alpha; EIF2A; eIF-2-alpha; eIF-2A; eIF-2alpha]

Human, Mouse, Rat

315

Two 96-Well Plates

Store at 4 degree C.

Reactivity Note: Human: S18, Mouse: S20, Rat: S19. Detection Method: Colorimetric 450 nm

Cell Based ELISA Kit

Background/Introduction: The Colorimetric Cell-Based ELISA Kit allows for the detection of various target proteins and the effects that certain stimulation conditions have on target protein expression in different cell lines. Qualitative determination of target protein concentration is achieved by an indirect ELISA format. In essence, the target protein is captured by target-specific primary (1 degree) antibodies while the HRP-conjugated secondary (2 degree) antibodies bind the Fc region of the 1 degree antibody. Through this binding, the HRP enzyme conjugated to the 2 degree antibody can catalyze a colorimetric reaction upon substrate addition. Due to the qualitative nature of the Cell-Based ELISA, multiple normalization methods are described: 1) a monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target absorbance values. 2) Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method is used to determine cell density. After staining, the results can be analyzed by normalizing the absorbance values to cell amounts, by which the plating difference can be adjusted. 3) If a phosphorylated target is being detected, an antibody against the non-phosphorylated counterpart will be provided for normalization purposes. The absorbance values obtained for the non-phosphorylated target can be used to normalize the absorbance values for the phosphorylated target.

4758256

NP_004085.1

NM_004094.4

P05198

36112

Activation Of The MRNA Upon Binding Of The Cap-binding Complex And EIFs, And Subsequent Binding To 43S Pathway (105970); BDNF Signaling Pathway (712093); Cap-dependent Translation Initiation Pathway (105967); Eukaryotic Translation Initiation Pathway (105966); Formation Of The Ternary Complex, And Subsequently, The 43S Complex Pathway (105969); GTP Hydrolysis And Joining Of The 60S Ribosomal Subunit Pathway (105973); Gene Expression Pathway (105937); Hepatitis C Pathway (173973); Hepatitis C Pathway (173907); Herpes Simplex Infection Pathway (377873)

The translation initiation factor EIF2 catalyzes the first regulated step of protein synthesis initiation, promoting the binding of the initiator tRNA to 40S ribosomal subunits. Binding occurs as a ternary complex of methionyl-tRNA, EIF2, and GTP. EIF2 is composed of 3 nonidentical subunits, the 36-kD EIF2-alpha subunit (EIF2S1), the 38-kD EIF2-beta subunit (EIF2S2; MIM 603908), and the 52-kD EIF2-gamma subunit (EIF2S3; MIM 300161). The rate of formation of the ternary complex is modulated by the phosphorylation state of EIF2-alpha (Ernst et al., 1987 [PubMed 2948954]).[supplied by OMIM, Feb 2010]

eIF2-alpha: a translation initiation factor that 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. Phosphorylated by at least 4 kinases: 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. Upregulated in some thyroid cancers and bronchiolo-alveolar adenocarcinomas; aberrant phosphorylation correlates with Alzheimer disease and Epstein-Barr virus infections. Protein type: RNA-binding; Translation; Translation initiation. Chromosomal Location of Human Ortholog: 14q23.3. Cellular Component: cytosol; eukaryotic translation initiation factor 2 complex; eukaryotic translation initiation factor 2B complex; extracellular exosome; membrane; nucleus; polysome; ribosome; stress granule; translation initiation ternary complex. Molecular Function: protein binding; ribosome binding; RNA binding; translation initiation factor activity. Biological Process: aging; cellular response to amino acid starvation; cellular response to heat; cellular response to UV; negative regulation of guanyl-nucleotide exchange factor activity; negative regulation of translational initiation in response to stress; PERK-mediated unfolded protein response; positive regulation of neuron death; positive regulation of type B pancreatic cell apoptotic process; protein autophosphorylation; response to manganese-induced endoplasmic reticulum stress; translational initiation; transmembrane transport

2 x96 Wells

535 USD