Detection of Human Phospho-eIF2 alpha (S52) by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HEK293T human embryonic kidney cell line untreated (-) or treated (+) with 100nM Calyculin A and 20 ng/mL Recombinant Human TNF‑ alpha (Catalog # 210-TA) for 10 minutes or 20 mJ/cm2 ultraviolet light (UV) followed by a 30 minute recovery. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Phospho-eIF2 alpha (S52) Monoclonal Antibody (Catalog # MAB39971) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-eIF2 alpha (S52) at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: eIF2 alpha
Eukaryotic translation initiation factor 2 alpha subunit (eIF2 alpha ) is a subunit of the eIF2 protein, an important regulator of translation initiation. Phosphorylation of eIF2 alpha on Ser 52 increases the affinity of eIF2 alpha for eIF2B, a guanine nucleotide exchange factor needed for the recycling of eIF2-GDP to eIF2-GTP. Reduction of eIF2-GTP levels leads to the suppression of the overall rate of protein synthesis. Heme-regulated inhibitor (HRI), ER-resident protein kinase PERK, dsRNA activated protein kinase PKR, and the homologue of the S. cerevisiae protein kinase GCN2 are all eIF2 alpha kinases.
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