Anti-Vitamin D Receptor antibody [EPR4552] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-Vitamin D Receptor antibody [EPR4552] - BSA and Azide free (AB239958)

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).

All lanes:

Immunoprecipitation - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (<a href='/en-us/products/primary-antibodies/vitamin-d-receptor-antibody-epr4552-chip-grade-ab109234'>ab109234</a>)

Predicted band size: 48 kDa

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• ChIP

Unknown

ChIP - 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).

• WB

Lab

Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - BSA and Azide free (AB239958)

This data was developed using the same antibody clone in a different buffer formulation (ab109234).

Lanes 1-4 : Merged signal (red and green). Green - ab109234 observed at 50 kDa. Red - loading control ab8245 observed at 37 kDa.

ab109234 Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade was shown to specifically react with Vitamin D Receptor in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265430 (knockout cell lysate ab257796) was used. Wild-type and Vitamin D Receptor knockout samples were subjected to SDS-PAGE. ab109234 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (<a href='/en-us/products/primary-antibodies/vitamin-d-receptor-antibody-epr4552-chip-grade-ab109234'>ab109234</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

Vitamin D Receptor knockout HeLa lysate at 20 µg

Lane 2:

Western blot - Human VDR (Vitamin D Receptor) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-vdr-vitamin-d-receptor-knockout-hela-cell-line-ab265430'>ab265430</a>)

Lane 3:

U-937 lysate at 20 µg

Lane 4:

SH-SY5Y lysate at 20 µg

Predicted band size: 48 kDa

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