MiliporeSigma
Anti-Sulf2, Clone 2B4
MABC584
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This product was used to investigate the expression of SULF2 across a number of different cell lines via western blot. Here we show how the antibody performed on lysate from MCF-7 cells.
Western Blot
Human MCF-7 Cell line lysate
Anti-SULF2 was used at 1 µg/mL in blocking buffer, overnight incubation at 4°C.
PBS 0.1% Tween20, 5% Blotting grade Non-fat dry milk, 1 hr incubation at RT
Secondary AB Sheep anti-Mouse was used at 1:50000 diluted in blocking buffer, incubated for 1.5 hrs at RT
N/A
ECL substrate SuperSignal West Femto Max Sensitivity
MCF-7 cells were treated with either scrambled or SULF2 siRNAs for knockdown of SULF2. After knockdown, lysate was collected from both groups using RIPA buffer with protease inhibitors (Halt) and protein concentration measured using BCA assay. 30 µg of lysate was loaded per lane of a pre-cast 10% bis-tris gel, which was run at 200 V using a MOPS SDS running buffer for 50 minutes. Transfer was done using 20X NuPage Transfer Buffer at 40 V for 5 hours at 4°C. Exposure time was 2 minute. Three bands were picked up by the antibody, ~80 kDa, ~95 kDa and ~125 kDa. All of which were not present in the knockdown (KD), and only the ~125 kDa (weakest) was predicted by the company. We are not sure what the other bands are.
Seems to work, but will require optimization for your cell line
Non-specific bands may be detected
It's an OK antibody but bear in mind you will need to optimize your system and cell line of choice.