6X His tag Mouse Monoclonal Antibody [8-F0]

Western blot analysis of 6X His tag on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M0812-3, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: C-terminal His-tagged recombinant protein
Lane 2: N-terminal His-tagged recombinant protein

His tag was immunoprecipitated in 2µg C terminal His Tag fusion protein lysate with M0812-3 at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using R1207-2 at 1/500 dilution. Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 60 mins at room temperature.

Lane 1: His Tag fusion protein lysate (input).
Lane 2: M0812-3 IP in His Tag fusion protein lysate.
Lane 3: Mouse IgG instead of M0812-3 in His Tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST