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Immunofluorescence analysis of eIF2a 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 eIF2a Mouse Monoclonal Antibody at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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, Non-human primate, Mouse, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human eIF-2-alpha.|
|Storage buffer||PBS, pH 7.4|
|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|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||1:500-1:1000|
* 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.
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.
Generation and characterization of an analog-sensitive PERK allele.
AHO0802 was used in western blot to study PERK signaling using an analog-sensitive PERK allele that accepts N(6)-alkylated ATP analogs
|Maas NL,Singh N,Diehl JA||Cancer biology & therapy (15:1106)||2014|
|Deficiency of ATP2C1, a Golgi ion pump, induces secretory pathway defects in endoplasmic reticulum (ER)-associated degradation and sensitivity to ER stress.||Ramos-Castañeda J,Park YN,Liu M,Hauser K,Rudolph H,Shull GE,Jonkman MF,Mori K,Ikeda S,Ogawa H,Arvan P||The Journal of biological chemistry (280:9467)||2005|
|Mouse||Not Cited||The PERK eukaryotic initiation factor 2 alpha kinase is required for the development of the skeletal system, postnatal growth, and the function and viability of the pancreas.||Zhang P,McGrath B,Li S,Frank A,Zambito F,Reinert J,Gannon M,Ma K,McNaughton K,Cavener DR||Molecular and cellular biology (22:3864)||2002|
|Mouse||Not Cited||VSV replication in neurons is inhibited by type I IFN at multiple stages of infection.||Trottier MD,Palian BM,Reiss CS||Virology (333:215)||2005|
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