SOT12 | Cytosolic Sulfotransferase 12

50 µg

Lyophilized antibody can be stored at -20°C for up to 3 years. Re-constituted antibody can be stored at 4°C for several days to weeks.
Once reconstituted, make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

1 :  1500 (WB)

37 kDa | 35 kDa

Immunoblotting of Arabidopsis thaliana total protein extracts with anti-SOT12 antibody (αSOT12). In the image above wild-type Col-0 is presented next to the SOT12-over-expressing mutant rcd1. The fragment of this image has been published in [https://elifesciences.org/articles/43284], Fig 6B. Leaves from mature plants were harvested and snap-frozen in liquid nitrogen. Frozen leaf material was ground to fine powder. Lysis buffer was added that contained 2% SDS, 20 mM Tris-HCl (pH 7.8), and x 100 protease inhibitor cocktail (Sigma-Aldrich #P9599). Upon addition of the buffer, the samples were vigorously mixed and then incubated 20 min at +37 ̊C. Supernatant was cleared by centrifugation for 5 min at room temperature on the desk-top mini-centrifuge at full speed. Protein concentration was measured using DC Protein Assay Kit II (Bio-Rad #5000112). Samples were equilibrated on equal protein basis. 100 μg of total protein was loaded per well. Protein was separated in precast 4-15 % gradient mini-gels (Bio-Rad #4561085DC) and transferred to PVDF membrane by wet transfer overnight at +4 ̊C and 130 mA, in 1 x Laemmli buffer supplemented with 20 % methanol. The membrane with transferred protein was blocked for 1 hour in TBS/Tween-20 (0.01 %) with 3 % BSA. αSOT12 antibody (diluted x 1 500 in 3 % BSA/TBS/Tween) was added, followed by overnight incubation at +4 ̊C. The membrane was washed 6 x 5 minutes with TBS/Tween. Secondary anti-rabbit HRP-conjugated antibody was added, diluted x 5 000 in 3 % BSA/TBS/Tween, and the membrane was incubated for 1 hour at room temperature. The membrane was washed 6 x 5 minutes with TBS/Tween. Immunoblotting signal was detected by chemiluminescence using the chemiluminescent detection reagent and the UVP BioSpectrum imaging system. The exposure time was 2 min.

Courtesy of Dr. Alexey Shapiguzov, Helsinki University, Finland

Plant and algal protein extraction buffer

Secondary antibodies