Western blot analysis of 6X His tag on C-terminal His-tagged recombinant protein lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (R1207-2, 1/2,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

His tag was immunoprecipitated in 2µg C terminal His Tag fusion protein lysate with R1207-2 at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using M0812-3 at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 60 mins at room temperature.

Lane 1: His Tag fusion protein lysate (input).
Lane 2: 0812-1 IP in His Tag fusion protein lysate.
Lane 3: R1207-2 IP in His Tag fusion protein lysate.
Lane 4: Rabbit IgG instead of R1207-2 in His Tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST