EIF1AX siRNA was transiently transfected into HEC‐1A cells. Cycloheximide (100 ng/mL) was added to the DMEM culture medium, and incubation was continued for the indicated time. The cell lysates were submitted for western blot analysis using mouse anti‐His mAb (Invitrogen), and western blot data were quantified using ImageJ software.
Mouse anti his mab
Manufactured by Thermo Fisher Scientific
The Mouse anti‐His mAb is a monoclonal antibody that specifically binds to the histidine (His) tag. It can be used to detect and purify recombinant proteins that have been engineered to contain a His-tag.
Automatically generated - may contain errors
2 protocols using mouse anti his mab
1
EIF1AX siRNA Transfection and Cycloheximide Assay
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EIF1AX siRNA was transiently transfected into HEC‐1A cells. Cycloheximide (100 ng/mL) was added to the DMEM culture medium, and incubation was continued for the indicated time. The cell lysates were submitted for western blot analysis using mouse anti‐His mAb (Invitrogen), and western blot data were quantified using ImageJ software.
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EIF1AX siRNA was transiently transfected into HEC‐1A cells. Cycloheximide (100 ng/mL) was added to the DMEM culture medium, and incubation was continued for the indicated time. The cell lysates were submitted for western blot analysis using mouse anti‐His mAb (Invitrogen), and western blot data were quantified using ImageJ software.
2
Western Blot Analysis of Transfected Cells
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Transfected cells were lysed in buffer containing 50 mM HEPES, 150 mM KCl, 2 mM EDTA, 0.5% NP-40, and protease inhibitor cocktail. Protein concentration in the lysates was measured by bicinchoninic protein assay and the lysates (40 μg) were separated by SDS-PAGE. The proteins were transferred to polyvinylidene difluoride membranes and probed with anti-HA mAb (Covance), mouse anti-His mAb (Invitrogen) and anti-GAPDH mAb (Life Technologies) followed by goat anti-mouse HRP-conjugated second antibody (Sigma). The blots were visualized using luminescent substrate (Millipore) on iBright CL1000.
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