All lanes :

Lane 1 : MCF7 whole cell lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : K562 whole cell lysate
Lane 4 : Jurkat whole cell lysate
Lane 5 : U2OS whole cell lysate
Lane 6 : HepG2 whole cell lysate
Lane 7 : C2C12 whole cell lysate
Lane 8 : NIH3T3 whole cell lysate
Lane 9 : NRK whole cell lysate

Lysates/proteins at 80 µg per lane.

Secondary
All lanes : goat anti-mouse-HRP at 1/20000 dilution

Predicted band size: 50 kDa

Immunofluorescent analysis of eIF2B3 (green) showing staining in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an eIF2B3 monoclonal antibody (ab171093) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Immunofluorescent analysis of eIF2B3 (green) showing staining in the cytoplasm of COS7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an eIF2B3 monoclonal antibody (ab171093) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

ab171093 staining eIF2B3 in the cytoplasm of Human uterus tissue (right) compared with a negative control in the absence of primary antibody (left) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

ab171093 staining eIF2B3 in the cytoplasm of Human breast tissue (right) compared with a negative control in the absence of primary antibody (left) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissue sections were incubated with the primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-mouse was used as the secondary antibody, followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Immunoprecipitation of U2OS cells labeling eIF2B3 with ab171093 at 3µg per 500µg lysate.