Anti-PDGFR beta antibody [42G12]

• WB

Lab

Western blot - Anti-PDGFR beta antibody [42G12] (AB69506)

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.

All lanes:

Western blot - Anti-PDGFR beta antibody [42G12] (ab69506) at 1/1000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysate at 30 µg

Lane 2:

PDGFRB knockout SH-SY5Y cell lysate at 30 µg

Lane 2:

Western blot - Human PDGFRB knockout SH-SY5Y cell line (<a href='/en-us/products/cell-lines/human-pdgfrb-knockout-sh-sy5y-cell-line-ab273749'>ab273749</a>)

Lane 3:

Human Skeletal Muscle tissue lysate at 30 µg

Lane 4:

HeLa cell lysate at 30 µg

Predicted band size: 123 kDa

Observed band size: 170 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR beta antibody [42G12] (AB69506)

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.

• ELISA

Lab

ELISA - Anti-PDGFR beta antibody [42G12] (AB69506)

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)

• Flow Cyt

Unknown

Flow Cytometry - Anti-PDGFR beta antibody [42G12] (AB69506)

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.