Anti-eIF2B3 antibody [1H3]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2B3 antibody [1H3] (AB171093)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2B3 antibody [1H3] (AB171093)

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-eIF2B3 antibody [1H3] (AB171093)

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 4°C 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-eIF2B3 antibody [1H3] (AB171093)

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 4°C 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.

• IP

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Immunoprecipitation - Anti-eIF2B3 antibody [1H3] (AB171093)

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

All lanes:

Immunoprecipitation - Anti-eIF2B3 antibody [1H3] (ab171093)

Predicted band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-eIF2B3 antibody [1H3] (AB171093)

All lanes:

Western blot - Anti-eIF2B3 antibody [1H3] (ab171093) at 1/1000 dilution

Lane 1:

MCF7 whole cell lysate at 80 µg

Lane 2:

HeLa whole cell lysate at 80 µg

Lane 3:

K562 whole cell lysate at 80 µg

Lane 4:

Jurkat whole cell lysate at 80 µg

Lane 5:

U2OS whole cell lysate at 80 µg

Lane 6:

HepG2 whole cell lysate at 80 µg

Lane 7:

C2C12 whole cell lysate at 80 µg

Lane 8:

NIH3T3 whole cell lysate at 80 µg

Lane 9:

NRK whole cell lysate at 80 µg

Secondary

All lanes:

goat anti-mouse-HRP at 1/20000 dilution

Predicted band size: 50 kDa

false