Recombinant Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

Lanes 1 - 4: Merged signal (red and green). Green - ab32570 observed at 170 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab32570 was shown to react with PDGFRB in wild-type SH-SY5Y cells in Western blot with weakened 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 fluorescent western blot (TBS-based) blocking solution before incubation with ab32570 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging. The weak signal observed in PDGFRB knockout cell line should be PDGFRA.

ab32570 staining PDGFR beta in human lung cancer tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Shows positive staining on stromal cells. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody (1/500). An HRP-conjugated Goat anti-rabbit IgG (ready to use) was used as the secondary antibody. Counter stained with Hematoxylin.

Immunofluorescence analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells stimulated with PDGF, staining PDGFR beta with unpurified ab32570.

Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling PDGFR beta (red) with ab32570 at a 1/20 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies. 

ELISA showing primary antibody ab32570 binding to the antigen Human PDGFR alpha protein and Human PDGFR beta protein. 

Primary antibody concentration ranges from 0 - 1000 ng/mL. 

Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) was used as a secondary antibody at 1/2500 dilution. 

Immunohistochemical staining of paraffin embedded human spleen with purified ab32570 at a working dilution of 1/50. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Immunofluorescence staining of NIH/3T3 (Mouse embryo fibroblast cell line) cells with purified ab32570 at a working dilution of 1 in 100, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor^® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor^® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.

The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab32570 was used at a dilution of 1/200 followed by an Alexa Fluor^® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

ab32570 (purified) at 1/20 immunoprecipitating PDGFR beta in NIH/3T3 (Mouse embryo fibroblast cell line) (Lane 1 and 2). Lane 3 - PBS.

For western blotting a HRP-conjugated anti-rabbit IgG specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

ab32570 staining PDGFR beta in human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Shows positive staining on stromal cells. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody (1/500). An HRP-conjugated Goat anti-rabbit IgG (ready to use) was used as the secondary antibody. Counter stained with Hematoxylin.