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Western blot analysis of 6X His tag on different lysates with Mouse anti-6X His tag antibody (HA601079) at different dilutions.
Lane 1/2: C-terminal His-tag fusion protein lysates
Lane 3/4: Mid His-tag fusion protein lysates
Lysates/proteins at 50 ng/Lane.
Observed band size: 30/40 kDa
Exposure time: 30 seconds;
12% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601079) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
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His-tag was immunoprecipitated in 2µg C-terminal His-tag fusion protein lysate with HA601079. Western blot was performed from the immunoprecipitate using HA601079 at 1/2,000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 60 mins at room temperature.
Lane 1: C-terminal His-tag fusion protein lysate (input).
Lane 2: HA601079 IP in C-terminal His-tag fusion protein lysate.
Lane 3: Mouse IgG instead of HA601079 IP in C-terminal His-tag fusion protein lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
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