EIF5B Antibody / Eukaryotic translation initiation factor 5B Antibody (FY13567)

EIF5B Antibody by IF in A549 cells. Immunofluorescence using Eukaryotic translation initiation factor 5B Antibody demonstrates widespread green cytoplasmic staining in A549 cells, surrounding nuclei counterstained blue with DAPI. This distribution is consistent with the cytoplasmic role of EIF5B in translation initiation. Following enzyme antigen retrieval for 15 minutes and blocking with 10% goat serum, cells were incubated with primary antibody at 5 ug/mL overnight at 4oC. Fluoro488-conjugated goat anti-rabbit IgG was applied at 1:500 for 30 minutes at 37oC.

EIF5B Antibody by IHC in human stomach carcinoma. Immunohistochemistry of paraffin-embedded human stomach carcinoma using Eukaryotic translation initiation factor 5B Antibody reveals strong brown cytoplasmic staining within carcinoma cell clusters. Sections underwent heat-mediated antigen retrieval in EDTA buffer at pH 8.0 and blocking with 10% goat serum. Primary antibody was applied at 2 ug/mL overnight at 4oC, followed by HRP-conjugated secondary antibody and DAB chromogen. Hematoxylin counterstaining highlights tissue architecture and nuclei.

EIF5B Antibody by WB in human A549. Western blot analysis using Eukaryotic translation initiation factor 5B Antibody detects an immunoreactive band near 200 kDa in human A549 (lane 1); human HepG2 (lane 2); human Jurkat (lane 3); human K562 (lane 4); rat RH35 (lane 5); mouse NIH/3T3 (lane 6) whole-cell lysates. The calculated molecular weight is approximately 139 kDa. The band migrates well above the calculated mass, and the cause of the difference has not been established. Reduced lysates containing 30 ug protein per lane were resolved by 8% SDS-PAGE and transferred to nitrocellulose. Following blocking with 5% nonfat milk, the membrane was incubated with primary antibody at 0.5 ug/mL overnight at 4oC and developed with an HRP-conjugated secondary antibody and chemiluminescence.

EIF5B Antibody by IHC in human stomach. Immunohistochemistry of paraffin-embedded human stomach using Eukaryotic translation initiation factor 5B Antibody reveals brown cytoplasmic staining in gastric glandular epithelium. Sections underwent heat-mediated antigen retrieval in EDTA buffer at pH 8.0 and blocking with 10% goat serum. Primary antibody was applied at 2 ug/mL overnight at 4oC, followed by HRP-conjugated secondary antibody and DAB chromogen. Hematoxylin counterstaining highlights tissue architecture and nuclei.

EIF5B Antibody by IHC in mouse brain. Immunohistochemistry of paraffin-embedded mouse brain using Eukaryotic translation initiation factor 5B Antibody reveals light brown cytoplasmic immunoreactivity in neuronal cell bodies. Sections underwent heat-mediated antigen retrieval in EDTA buffer at pH 8.0 and blocking with 10% goat serum. Primary antibody was applied at 2 ug/mL overnight at 4oC, followed by HRP-conjugated secondary antibody and DAB chromogen. Hematoxylin counterstaining highlights tissue architecture and nuclei.

EIF5B Antibody by IHC in rat brain. Immunohistochemistry of paraffin-embedded rat brain using Eukaryotic translation initiation factor 5B Antibody reveals brown staining within neuronal somata and neuropil. Sections underwent heat-mediated antigen retrieval in EDTA buffer at pH 8.0 and blocking with 10% goat serum. Primary antibody was applied at 2 ug/mL overnight at 4oC, followed by HRP-conjugated secondary antibody and DAB chromogen. Hematoxylin counterstaining highlights tissue architecture and nuclei.

EIF5B Antibody by FACS in Jurkat cells. Intracellular flow cytometry using Eukaryotic translation initiation factor 5B Antibody demonstrates a fluorescence shift in fixed and permeabilized Jurkat cells relative to the rabbit IgG isotype control. The antibody-stained population is shown in blue, isotype control in green, and unstained control in red. Cells were fixed with 4% paraformaldehyde, permeabilized, blocked with 10% goat serum, and incubated with 1 ug primary antibody per 1 x 10^6 cells for 30 minutes at 20oC. Fluorescent secondary antibody was used for detection.

EIF5B Antibody by IP in human HepG2 cells. Immunoprecipitation using Eukaryotic translation initiation factor 5B Antibody was evaluated by Western blot of HepG2 whole-cell lysate. Lane 1 contains 30 ug input lysate, lane 2 is the rabbit IgG immunoprecipitation control, and lane 3 contains material precipitated with 2 ug primary antibody from 500 ug lysate. A high-molecular-weight band near 200 kDa is evident in the input and immunoprecipitated material, above the approximately 139 kDa calculated EIF5B mass. The membrane was probed with primary antibody at 0.5 ug/mL and detected using an HRP-conjugated secondary antibody and chemiluminescence.