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PDGFR beta Recombinant Rabbit Monoclonal Antibody [SY10-08] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750082

PDGFR beta Recombinant Rabbit Monoclonal Antibody [SY10-08] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IP

  • FC

  • IF-Tissue

  • IF-Cell

  • IHC-Fr

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

PDGFR beta Recombinant Rabbit Monoclonal Antibody [SY10-08] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human PDGFR beta aa 961-1,106 / 1,106.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP, FC, IF-Tissue, IF-Cell, IHC-Fr

Molecular Weight

Predicted band size: 124 kDa

Positive Control

SH-SY5Y cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, SH-SY5Y, human spleen tissue, mouse brain tissue, mouse spleen tissue, rat spleen tissue.

Conjugation

unconjugated

Clone Number

SY10-08

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:1,000

  • IF-Tissue

  • 1:200

  • IF-Cell

  • 1:100

  • IHC-Fr

  • 1:200

Target

Function

Platelet-derived growth factor (PDGF) is a mitogen for mesenchyme- and glia-derived cells. PDGF consists of two chains, A and B, which dimerize to form functionally distinct isoforms, PGDF-AA, PDGF-AB and PDGF-BB. These three isoforms bind with different affinities to two receptor types, PDGFR-α and –β, which are endowed with protein tyrosine kinase domains. PDGFR-α can bind to both A and B subunits of PDGF, while PDGFR-β can only bind the B subunit. Ligand binding promotes either homo- or heterodimerization of the PDGF receptors in a specific manner. PDGF-AA induces the dimerization of two α receptors, PDGF-AB induces dimerization of αα and αβ and PDGF-BB induces the formation of three types of dimers, αα, αβ and ββ. Translocation of the PDGFR-∫ gene with the Tel gene is linked to chronic myelomonocytic leukemia (CMML), a myelodysplastic syndrome, and demonstrates the oncogenic potential of the PDGF receptors.

Background References

1. Cui Z et al. Endothelial PDGF-BB/PDGFR-beta signaling promotes osteoarthritis by enhancing angiogenesis-dependent abnormal subchondral bone formation. Bone Res. 2022 Aug

2. Hu G et al. PDGFR-beta(+) fibroblasts deteriorate survival in human solid tumors: a meta-analysis. Aging (Albany NY). 2021 May

Sequence Similarity

Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.

Post-translational Modification

Autophosphorylated on tyrosine residues upon ligand binding. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit. Phosphorylation at Tyr-579, and to a lesser degree, at Tyr-581, is important for interaction with SRC family kinases. Phosphorylation at Tyr-740 and Tyr-751 is important for interaction with PIK3R1. Phosphorylation at Tyr-751 is important for interaction with NCK1. Phosphorylation at Tyr-771 and Tyr-857 is important for interaction with RASA1/GAP. Phosphorylation at Tyr-857 is important for efficient phosphorylation of PLCG1 and PTPN11, resulting in increased phosphorylation of AKT1, MAPK1/ERK2 and/or MAPK3/ERK1, PDCD6IP/ALIX and STAM, and in increased cell proliferation. Phosphorylation at Tyr-1009 is important for interaction with PTPN11. Phosphorylation at Tyr-1009 and Tyr-1021 is important for interaction with PLCG1. Phosphorylation at Tyr-1021 is important for interaction with CBL; PLCG1 and CBL compete for the same binding site. Dephosphorylated by PTPRJ at Tyr-751, Tyr-857, Tyr-1009 and Tyr-1021. Dephosphorylated by PTPN2 at Tyr-579 and Tyr-1021.; N-glycosylated.; Ubiquitinated. After autophosphorylation, the receptor is polyubiquitinated, leading to its degradation.

Subcellular Location

Cell membrane, Cytoplasmic vesicle, Lysosome lumen.

UNIPROT

SwissProt: P05622 Mouse

SwissProt: P09619 Human

SwissProt: Q05030 Rat

Synonyms

Beta platelet derived growth factor receptor antibody

Beta-type platelet-derived growth factor receptor antibody

CD 140B antibody

CD140 antigen-like family member B antibody

CD140b antibody

CD140b antigen antibody

IBGC4 antibody

IMF1 antibody

JTK12 antibody

OTTHUMP00000160528 antibody

Expand

Images

  • Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/200 dilution.<br /><br /><span style="font-weight: bold;">The section was not undergone antigen retrieval.</span> The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-PDGFR beta antibody (HA750082) at 1/200 dilution.

    The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750082, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/200 dilution.<br /><br /><span style="font-weight: bold;">The section was not undergone antigen retrieval.</span> The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-PDGFR beta antibody (HA750082) at 1/200 dilution.

    The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750082, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Western blot analysis of PDGFR beta on different lysates with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 2: C6 cell lysate (20 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 4: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 124 kDa<br />Observed band size: 190 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PDGFR beta on different lysates with Rabbit anti-PDGFR beta antibody (HA750082) at 1/1,000 dilution.

    Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 2: C6 cell lysate (20 µg/Lane)
    Lane 3: Mouse brain tissue lysate (40 µg/Lane)
    Lane 4: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 124 kDa
    Observed band size: 190 kDa

    Exposure time: 6 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750082) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of SH-SY5Y cells labeling PDGFR beta with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SH-SY5Y cells labeling PDGFR beta with Rabbit anti-PDGFR beta antibody (HA750082) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PDGFR beta antibody (HA750082) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-PDGFR beta antibody (HA750082) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750082) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-PDGFR beta antibody (HA750082) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750082) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-PDGFR beta antibody (HA750082) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750082) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-PDGFR beta antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-PDGFR beta antibody (HA750082) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750082) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of SH-SY5Y cells labeling PDGFR beta.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SH-SY5Y cells labeling PDGFR beta.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA750082, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Mouse<br />Tissue: Spleen<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br />Primary antibody: <a href="/products/HA750082" style="font-weight: bold;text-decoration: underline;">HA750082</a>, 1/200, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Mouse
    Tissue: Spleen
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
    Primary antibody: HA750082, 1/200, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"