The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/200 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
Use at 2-5 µg/mg of lysate.
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.
Belongs to the eIF-2-alpha family. Contains 1 S1 motif domain.
Substrate 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.
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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma (left) and mouse teratoma (right) tissues labelling EIF2S1 with ab70542 at 1/1000 (0.2µg/ml) and 1/200 (1µg/ml). Detection: DAB.
Western blot - EIF2S1 antibody (ab70542)
All lanes : Anti-EIF2S1 antibody (ab70542) at 1/5000 dilution
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg Lane 5 : Whole cell lysate from NIH3T3 cells at 50 µg
Developed using the ECL technique.
Predicted band size: 36 kDa Observed band size: 36 kDa
Exposure time: 30 seconds
Immunoprecipitation - EIF2S1 antibody (ab70542)
1mg of whole cell lysate was immunoprecipitated with ab70542 at 3ug/mg of lysate (lane 1) or a control rabbit IgG (lane 2). For the subsequent western blot, 20% of the immunoprecipitated was loaded per lane, and ab70542 was used at 1ug/ml.
Xia X et al. Arginine Supplementation Recovered the IFN-?-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2a Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells. Mol Cells39:410-7 (2016).
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