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Immunofluorescence analysis of EIF2-alpha was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with EIF2-alpha Rabbit Polyclonal Antibody (AHO1182) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Rat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS, pH 7.2, with 1% BSA|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Western Blot (WB)||0.5-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
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]).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Measurement of the unfolded protein response to investigate its role in adipogenesis and obesity.
AHO1182 was used in western blot to discuss the role of the unfolded protein response in obesity pathogenesis
|Han J,Kaufman RJ||Methods in enzymology (538:135)||2014|
|Rat||Not Cited||Impaired dexamethasone-mediated induction of tryptophan 2,3-dioxygenase in heme-deficient rat hepatocytes: translational control by a hepatic eIF2alpha kinase, the heme-regulated inhibitor.||Liao M,Pabarcus MK,Wang Y,Hefner C,Maltby DA,Medzihradszky KF,Salas-Castillo SP,Yan J,Maher JJ,Correia MA||The Journal of pharmacology and experimental therapeutics (323:979)||2007|
EIF-2; eIF-2-alpha; EIF-2A; EIF-2alpha; EIF2; EIF2A; EIF2S1; eukaryotic translation initiation factor 2 subunit 1; eukaryotic translation initiation factor 2 subunit alpha; eukaryotic translation initiation factor 2, subunit 1 alpha, 35kDa; eukaryotic translation initiation factor 2A
0910001O23Rik; 2410026C18Rik; 35kDa; EIF-2; EIF-2A; EIF-2alpha; EIF2; EIF2A; eIF2alpha; EIF2S1