Key features and details
- Rabbit polyclonal to Heme-regulated inhibitor
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Heme-regulated inhibitor antibody
See all Heme-regulated inhibitor primary antibodies
DescriptionRabbit polyclonal to Heme-regulated inhibitor
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Cynomolgus monkey
- HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Previously labelled as EIF2AK1.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab28530 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).|
FunctionInhibits protein synthesis at the translation initiation level, in response to various stress conditions, including oxidative stress, heme deficiency, osmotic shock and heat shock. Exerts its function through the phosphorylation of EIF2S1 at 'Ser-48' and 'Ser-51', thus preventing its recycling. Binds hemin forming a 1:1 complex through a cysteine thiolate and histidine nitrogenous coordination. This binding occurs with moderate affinity, allowing it to sense the heme concentration within the cell. Thanks to this unique heme-sensing capacity, plays a crucial role to shut off protein synthesis during acute heme-deficient conditions. In red blood cells (RBCs), controls hemoglobin synthesis ensuring a coordinated regulation of the synthesis of its heme and globin moieties. Thus plays an essential protective role for RBC survival in anemias of iron deficiency. Similarly, in hepatocytes, involved in heme-mediated translational control of CYP2B and CYP3A and possibly other hepatic P450 cytochromes. May also contain ER stress during acute heme-deficient conditions.
Tissue specificityExpressed predominantly in erythroid cells. At much lower levels, expressed in hepatocytes (at protein level).
Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
Contains 2 HRM (heme regulatory motif) repeats.
Contains 1 protein kinase domain.
modificationsActivated by autophosphorylation; phosphorylated predominantly on serine and threonine residues, but also on tyrosine residues. Autophosphorylation at Thr-488 is required for kinase activation. The active autophosphorylated form apparently is largely refractory to cellular heme fluctuations.
- Information by UniProt
- E2AK1_HUMAN antibody
- EC 184.108.40.206 antibody
- Eif2ak1 antibody
Anti-Heme-regulated inhibitor antibody (ab28530) at 1 µg/ml + HepG2 whole cell lysate (ab7900) at 20 µg
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Additional bands at: 150 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab28530 stained human HepG2 cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab28530, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
IHC image of ab28530 staining Heme-regulated inhibitor in Human liver cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28530, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab28530 has been referenced in 1 publication.
- Vincent HA et al. Mechanism of Protein Kinase R Inhibition by Human Cytomegalovirus pTRS1. J Virol 91:N/A (2017). PubMed: 27974558