Anti-PDGFR beta antibody [42G12] (ab69506)

Immunohistochemistry analysis of human spleen tissue labeling PDGFR beta [42G12] with ab69506 at 20µg/ml.

This image was generated using the ascites version of the product.

Lanes 1 - 4: Merged signal (red and green). Green - ab69506 observed at 170 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab69506 was shown to react with PDGFR beta in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRB knockout cell line ab273749 (knockout cell lysate ab275523). Wild-type SH-SY5Y and PDGFRB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab69506 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

Immunohistochemistry analysis of human placenta tissue labeling PDGFR beta [42G12] with ab69506 at at 20µg/ml.

This image was generated using the ascites version of the product.

Overlay histogram showing NIH 3T3 cells stained with ab69506 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab69506, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This image was generated using the ascites version of the product.

ELISA using ab69506 at varying antibody concentrations (1000-0ng/ml). The antigens used were Human PDGFR alpha protein, Human PDGFR beta protein at a concentration of 1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Mouse IgG(H+L) was used as the secondary antibody with a dilution of 1:1000. The substrate solution used was p-nitrophenyl phosphate(PNPP)