Anti-PDGFR alpha antibody [EPR5480]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PDGFR alpha with ab134123 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stromal cells in human colon (PMID : 23429582). The section was incubated with ab134123 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Immunohistochemical analysis of paraffin-embedded Human colonic carcinoma tissue labeling PDGFR alpha with ab134123 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stromal cells in human colonic carcinoma. The section was incubated with ab134123 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Flow cytometry analysis of SH-SY5Y(Human neuroblastoma epithelial cell) cells labelling PDGFR alpha with ab134123 at 1/20 dilution (14.65 © : g/ml) (Red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor� 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue).

• I-ELISA

Supplier Data

Indirect ELISA - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

ELISA analysis of Human PDGFR alpha recombinant protein at 250 ng/mL with ab134123. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Flow cytometry analysis of L6(Rat skeletal muscle myoblast) cells labelling PDGFR alpha with ab134123 at 1/20 dilution (14.65 © : g/ml) (Red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor� 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue).

• WB

Supplier Data

Western blot - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Blocking/Dilution buffer : 5% NFDM/TBST. We recommend using higher sensitivity ECL to improve results. ab134123 can be a good alternative when testing samples with low level of PDGFR alpha which detects stronger signal than ab203491 in western blot.

Lanes 1 - 8:

Western blot - Anti-PDGFR alpha antibody [EPR22059-270] (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-antibody-epr22059-270-ab203491'>ab203491</a>) at 1/1000 dilution

Lanes 9 - 12:

Western blot - Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution

Lanes 1, 5 and 9:

SH-SY5Y

Lanes 2, 6 and 10:

Human brain tissue lysate at 20 µg

Lanes 3, 7 and 11:

Human heart tissue lysate at 20 µg

Lanes 4, 8 and 12:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

Western blot at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 150 kDa

false

Exposure time: 80s

• WB

Unknown

Western blot - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

All lanes:

Western blot - Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution

Lane 1:

Human skeletal muscle lysate at 10 µg

Lane 2:

Human spleen lysate at 10 µg

Lane 3:

Human fetal brain lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 190 kDa

false

• WB

Lab

Western blot - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Lanes 1 - 2 : Merged signal (red and green). Green - ab134123 observed at 180 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab134123 was shown to react with PDGFR alpha in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRA knockout cell line ab275335 (knockout cell lysate ab275522). Wild-type SH-SY5Y and PDGFRA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab134123 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 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.

All lanes:

Western blot - Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysate at 40 µg

Lane 2:

PDGFRA knockout SH-SY5Y cell lysate at 40 µg

Lane 2:

Western blot - Human PDGFRA knockout SH-SY5Y cell line (<a href='/en-us/products/cell-lines/human-pdgfra-knockout-sh-sy5y-cell-line-ab275335'>ab275335</a>)

Predicted band size: 122 kDa

Observed band size: 180 kDa

false

• WB

Unknown

Western blot - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

All lanes:

Western blot - Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution

Lane 1:

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

Lane 2:

C2C12 (mouse myoblast cell line) cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 190 kDa

false

• WB

Supplier Data

Western blot - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Blocking/Dilution buffer : 5% NFDM/TBST. We recommend using higher sensitivity ECL to improve results. ab134123 can be a good alternative when testing samples with low level of PDGFR alpha which detects stronger signal than ab203491 in western blot.

Lanes 1 - 10:

Western blot - Anti-PDGFR alpha antibody [EPR22059-270] (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-antibody-epr22059-270-ab203491'>ab203491</a>) at 1/1000 dilution

Lanes 11 - 15:

Western blot - Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution

Lanes 1, 6 and 11:

Rat heart tissue lysate at 20 µg

Lanes 2, 7 and 12:

Rat lung tissue lysate at 20 µg

Lanes 3, 8 and 13:

Mouse brain tissue lysate at 20 µg

Lanes 4, 9 and 14:

Mouse heart tissue lysate at 20 µg

Lanes 5, 10 and 15:

Mouse lung tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 150 kDa

false

Exposure time: 80s

• WB

Supplier Data

Western blot - Anti-PDGFR alpha antibody [EPR5480] (AB134123)

Blocking/dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution

Lane 1:

Rat heart tissue lysate at 20 µg

Lane 2:

Rat lung tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

Mouse lung tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 122 kDa

Observed band size: 150 kDa

false

Exposure time: 80s