Recombinant Anti-Vitamin D Receptor antibody [EPR4552] - BSA and Azide free (ab239958)

Chromatin was prepared from T-47D cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 2 µg of ab223850 (blue), and 20 µL of protein A/G sepharose beads slurry (10 µL of sepharose A beads + 10 µL of sepharose G beads). 5 μg of rabbit normal IgG was added to the beads as a control sample (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR Green chemistry) with primers to c-Fos.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109234).

Lane 1 (input): T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate, 10 μg
Lane 2(+): T-47D whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109234 in T-47D whole cell lysate

Ab109234 immunoprecipitating Vitamin D receptor in T-47D whole cell lysates. Capture antibody was used at a 1/30 dilution (2 μg in 0.35 mg lysates). For western blotting, primary antibody was used as ab109234 at 1/1000 dilution (0.62 μg/mL). VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109234).