Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal

• WB

Unknown

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution

All lanes:

Human fetal brain tissue lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Observed band size: 175 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling PDGFR alpha + PDGFR beta with ab32570 at a concentration of 0.5µg/ml.

The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5. ab32570 anti-PDGFR alpha + PDGFR beta was incubated at 37°C for 16min.

Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling PDGFR alpha + PDGFR beta with ab32570 at a concentration of 0.1µg/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.

ab32570 anti-PDGFR alpha + PDGFR beta antibody [Y92] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + 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.

• ELISA

Supplier Data

ELISA - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (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.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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

Image from Miyata M et al. J Biol Chem. 2009 Sep 4;284(36):24595-609. Epub 2009 Jul 9. Fig 1.; doi: 10.1074/jbc.M109.016436; September 4 2009 The Journal of Biological Chemistry 284 24595-24609.

• IP

Unknown

Immunoprecipitation - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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.

All lanes:

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

Observed band size: 175 kDa

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• WB

Unknown

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/10000 dilution

All lanes:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Observed band size: 175 kDa

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• WB

Lab

Western blot - 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.

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysate at 30 µg

Lane 2:

PDGFR beta 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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• WB

Unknown

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/50000 dilution

All lanes:

NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Observed band size: 175 kDa

false

• WB

Unknown

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/10000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Observed band size: 175 kDa

false

• WB

Lab

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Different batches of ab32570 were tested on Rat brain lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 175 kDa.

All lanes:

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

false

• WB

Lab

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/1000 dilution

Lane 1:

Western blot - Recombinant human PDGFR beta protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pdgfr-beta-protein-ab60833'>ab60833</a>) at 0.1 µg

Lane 2:

Western blot - Recombinant human PDGFR alpha protein (<a href='/en-us/products/proteins-peptides/recombinant-human-pdgfr-alpha-protein-ab84797'>ab84797</a>) at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

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Exposure time: 3s