Anti-SBPase | Sedoheptulose-1,7-bis phosphatase

Janna Setzkorn
Whole cell samples of Chlamydomonas reinhardtii corresponding to 2 µg chlorophyll were separated on 12 % SDS-PAGE (Laemmli-type) and blotted for 1.5 h on a PVDF membrane. After blocking (3% milk in TBS-T) for 1h at RT, blots were incubated in the primary antibody at a dilution of 1:1000 overnight at 4 °C with agitation. The primary antibody, diluted in 3% milk in TBS-T could only be reused once. The secondary HRP-linked anti-rabbit antibody was applied at a dilution of 1:1000 for about 1h at RT. However, using ECL detection, individual bands of SBPase could only be detected in samples from Chlamydomonas lines overexpressing SBPase, but not in the wild-type control.	2024-02-12 @ 10:26:43
Dorothée Klein
We analyzed whole cell samples of Chlamydomonas reinhardtii using 10 µg total protein per lane. After WB transfer, we used an AB dilution of 1:3000. The product worked good but I recommend to increase the amount of antibody or the amount of total protein used to get a better signal. We obtained signals for SBPase at approximately 42 kDa.	2023-03-23 @ 16:48:22