Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 293T (human embryonic kidney epithelial cell) transfected with a human wild-type PDGFRA expression vector containing a Myc tag (left); human PDGFRA (D842V) expression vector containing a Myc tag (right). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Cell labelling of PDGFRA (D842V) with ab313771 at 1/50 dilution. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used at 1/5000 dilution. No cross-reactivity with human wild-type PDGFRA.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A, D) human endometrial tissue; (B, E) human placenta; (C, F) human colon carcinoma.

A-C : Sections were incubated with anti-PDGFRA [EPR22059-270] (ab203491; 1 : 2000 [0.28µg/mL]) for 30 mins at room temperature. Positive staining on human endometrium (image A), placenta (image B), and colon carcinoma (image C). D-F : Sections were incubated with anti-PDGFRA D842V [EPR26304-280] (ab313771; 1 : 4000 [0.13µg/mL]) for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument counterstained with Hematoxylin. No staining on human endometrium (image D), placenta (image E), or colon carcinoma (image F). No cross-reactivity with human wild-type PDGFRA.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) labelling PDGFRA (D842V) with ab313771 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Cells were counterstained with anti-Myc tag Mouse mab [9E10] (Alexa Fluor^® 594) at 1/100 dilution (Red). The Nuclear counterstain is DAPI (Blue).

Confocal image showing cytoplasmic and membrane staining in 293T overexpressing human PDGFRA (D842V). No staining in 293T cells overexpressing human wild-type PDGFRA. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded 293T (human embryonic kidney epithelial cell) transfected with (A & D) human PDGFRA (D842V) expression vector containing a His tag; (B & E) human wild-type PDGFRA expression vector containing a His tag; (C & F) empty vector containing a His tag. A-C : whole cell pellet labelling of human wild-type and mutated PDGFRA with ab203491 at 1/2000 dilution (0.28 µg/ml). D-F : whole cell pellet labelling of human PDGFRA (D842V) with ab313771 at 1/4000 dilution (0.13 µg/ml). Sections were incubated with ab203491or ab313771 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin This antibody recognizes human PDGFR (D842V) but not human wild-type PDGFRA. Positive staining on 293T overexpressing human PDGFR (D842V) (image D). No staining on 293T overexpressing human wild-type PDGFRA (image E) or the empty vector control (image F). Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed 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 (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling PDGFR alpha (mutated D842V) with ab313771 at 1/4000 (0.13 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining was observed in all normal tissues tested. The section was incubated with ab313771 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 LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation. PDGFRA (D842V) was immunoprecipitated from 0.35 mg of 293T (human embryonic kidney epithelial cell) transfected with a human PDGFRA (D842V) expression vector containing a His tag using ab313771 at 1/30 dilution (2µg in 0.35mg whole cell lysate). Western blot was performed on the immunoprecipitate using ab313771 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730). Lanes 1-3 : 293T (human embryonic kidney epithelial cell) transfected with a human PDGFRA (D842V) expression vector containing a His tag Lane 1 : whole cell lysate at 2µg Lane 2 : IP with ab313771 Lane 3 : IP with Isotype control (ab172730) Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds

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Immunoprecipitation - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-mutated-d842v-antibody-epr26304-280-ab313771'>ab313771</a>) at 1/30 dilution

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293T cells transfected with a human PDGFRA (D842V) expression vector containing a His-tag whole cell lysate

Secondary

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Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 24s

• WB

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Western blot - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] - BSA and Azide free (AB313772)

This data was developed using ab313771, the same antibody clone in a different buffer formulation. This antibody recognizes human PDGFR (D842V) but not human wild-type PDGFRA. In western blot, anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution and anti-PDGFRA antibody (ab203491) staining at 1/1000 dilution. A-172 cells are negative for wild-type PDGFRA (PMID : 8425771).

All lanes:

Western blot - Anti-PDGFR alpha (mutated D842V) antibody [EPR26304-280] (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-mutated-d842v-antibody-epr26304-280-ab313771'>ab313771</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human wild-type PDGFRA expression vector containing a His-tag, whole cell lysate at 2 µg

Lane 3:

293T cells transfected with a human PDGFRA (D842V) expression vector containing a His-tag, whole cell lysate at 2 µg

Lane 4:

293T cells transfected with a human PDGFRA (D842V) expression vector containing a His-tag, whole cell lysate at 0.4 µg

Lane 5:

MG-63 (human osteosarcoma fibroblast) whole cell lysate at 20 µg

Lane 6:

A-204 (human muscle rhabdomyosarcoma cell) whole cell lysate at 20 µg

Lane 7:

A-172 (human brain glioblastoma ) whole cell lysate at 20 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 1/100000 dilution

Observed band size: 160 kDa

false

Exposure time: 180s