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, Not Applicable|
|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 and therapy (15:1106)||2014|
Endoplasmic reticulum stress induced by aqueous extracts of cigarette smoke in 3T3 cells activates the unfolded-protein-response-dependent PERK pathway of cell survival.
AHO0802 was used in western blot to investigate how cigarette smoke induces endoplasmic reticulum stress
|Hengstermann A,Müller T||Free radical biology and medicine (44:1097)||2008|
|Not Applicable||Not Cited||
Enhanced antiglioma activity of chimeric HCMV/HSV-1 oncolytic viruses.
AHO0802 was used in western blot to characterize herpes simplex virus-1 gamma(1)34.5 mutants
|Shah AC,Parker JN,Gillespie GY,Lakeman FD,Meleth S,Markert JM,Cassady KA||Gene therapy (14:1045)||2007|
|Not Applicable||Not Cited||
Human cytomegalovirus TRS1 and IRS1 gene products block the double-stranded-RNA-activated host protein shutoff response induced by herpes simplex virus type 1 infection.
AHO0802 was used in western blot to elucidate how human cytomegalovirus evades PKR-mediated protein shutoff and maintains late viral protein synthesis
|Cassady KA||Journal of virology (79:8707)||2005|
|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|