DNA Photolyase (E,coli)

Samples: Purified E.coli photolyase 8.4 ng (1), Purified E.coli photolyase 42 ng (2), E. coli crude lysate, 40 μg (3) were separated on 12.5 % SDS-PAGE and transferred to a PVDF membrane at 15V, overnight, followed by blocking with 5 % non-fat milk, washes and incubation with a primary antibodies at 1: 1000 dilution 1h/RT in TBS-T, washes and incubation with secondary antibodies HRP conjugated. Reaction was visualized by chemiluminescnce following manufacture's recommendations.

Important note: 

Immunoprecipitation was performed by using Dynabeads Magnetic system (Thermo Fisher).
Samples: E. coli AB1157 cell lysate (300μg) , primary antibody 2μg (1),   E. coli AB1157 cell lysate (300μg), antibody 10 μg (2), Positve control: E. coli Photolyase purified, 4.2 μg, antibody 10 μg (3), Negative control: No sample, antibody 10 μg (4). Western blot: Anti-Photolyase antibody used at 1 : 1000 dilution and as the second antibody, HRP-conjugated goat anti-rabbit IgG for IP (suitable for detection of immunprecipitation products) was used at 1: 1000 dilution 1 h/RT. Reaction was visualized by chemiluminescence, following manufacture's recommendations.