Product Name:
EIF2AK1-pS258
Product Number:
ab-pk876
Target Full Name: Eukaryotic translation initiation factor 2-alpha kinase 1
Target Alias: E2AK1; EC 2.7.11.1; EIF2AK1; HCR; HRI; Heme-controlled repressor; heme-regulated inhibitor; Hemin-sensitive initiation factor-2 alpha kinase
Product Type Specific: HRI protein kinase phosphosite-specific antibody
Antibody Code: PK876
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S258
Protein UniProt: Q9BQI3
Protein SigNET: EIF2AK1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human EIF2AK1
Antibody Immunogen Sequence: EVL(pS)DQE(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues E255 to E261. The effect of S258 phosphorylation is unknown. It appears to be the most commonly observed phosphosite in HRI from high throughput mass spectrometry studies (≥12 reports).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.75 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Rat | Mouse.
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Scientific Background: HRI is a protein-serine/threonine kinase in the Other Group and PEK family. It serves as an activator of the integrated stress response (ISR) required for adaptation to stress, such as heme deficiency, oxidative stress, osmotic shock, mitochondrial dysfunction and heat shock. It phosphorylates the alpha subunit of eukaryotic translation initiation factor 2 (EIF2S1/eIF-2-alpha) in response to various stress conditions, resulting in an inhibition of protein synthesis. It is synthesized in an inactive form that binds to the N-terminal domain of CDC37. It has to be associated with a multiprotein complex containing Hsp90, CDC37 and PPP5C for maturation and activation by autophosphorylation. The phosphatase PPP5C modulates this activation. It is a homodimer, non-covalently bound in the absence of hemin. Hemin inactivates EIF2AK1 by promoting the formation of a disulfide-linked homodimer. The binding of nitric oxide (NO) to the heme iron in the N-terminal heme-binding domain activates the kinase activity. The binding of carbon monoxide (CO) suppresses kinase activity. HRI was found to be down-regulated in the majority of ovarian cancers. However, in general in human cancers, it is up-regulated about 2.4-times the rate observed for most protein kinases. This description may include information annotated by UniProt and/or Google AI.