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

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.

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

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

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.

Immunohistochemistry (PFA perfusion fixed frozen sections) of rat kidney tissue labeling PDGFR beta with ab32570 at 1/100 dilution. Sections were fixed with Paraformaldehyde and permeabilised with 0.05% Tween 20. Antigen retrieval was heat mediated using ab64236, 100x Citrate Buffer pH 6.0. The secondary antibody was Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed antibody at 1/300.

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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.

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.

Human aortic valve: immunofluorescence labelling shows telocytes.

Frozen sections were sliced into a thickness of 6 μM, and then were post‐fixed with 4% paraformaldehyde dissolved in 0.1 M phosphate buffer (pH = 7.4) for at least 15 min. After washed with phosphate buffer for three times, sections were immersed in 10% goat serum for 1 hr. After that, sections were incubated overnight at 4°C with rat monoclonal anti‐CD34 (ab8158; Abcam, Cambridge, UK) and either rabbit monoclonal to Vimentin (ab92547; Abcam), rabbit monoclonal to PDGF Receptor‐beta (ab32570, 1:100; Abcam) or rabbit polyclonal anti‐C‐Kit (ab5506, 1:100; Abcam), both were with the dilution of 1:100 in phosphate buffer and permeabilized by 0.25% Triton X‐100 at the same time. On the second day, the sections were incubated to goat anti‐rabbit labelled with rhodamine secondary antibodies and goat anti‐rat labelled with FITC diluted 1:200 in phosphate buffer for 1 hr. The sections were stained with 4′,6‐diamidino‐2‐phenylindole (DAPI). 

Newborn rat pups received a single injection of GA (1 μmol/g) or vehicle into the cisterna magna. they were analysated analyzed at 14 and 30 days post injection (DPI) Characterization of EB positive cells was made by double labeling with ab32570 an antibody to β receptor of the platelet derived growth factor (PDGFRβ) at a dilution of 1/100.

Immunohistochemistry assays were performed on free-floating sections. Sections were washed with PBS; permeabilized 20 min with 0.05% Triton X-100 and incubated 30 min in blocking buffer (PBS containing 0.05% Triton X-100 and 5% bovine serum albumin (BSA)).

Representative images of the immunoreactivity for the pericyte marker PDGFRβ evidencing strong signal around some blood vessels (white arrows) and some middle-sized positive cells (arrowheads). At 30 DPI, there was a reduction in the PDGFRβ signal around blood vessels of GA-injected ratwhen compared to controls.

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.