Anti-PDGFR alpha 抗体 [EPR22059-270] (ab203491)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22059-270] to PDGFR alpha
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP, Indirect ELISA, IHC-Fr
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-PDGFR alpha antibody [EPR22059-270]
PDGFR alpha 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR22059-270] to PDGFR alpha -
由来種
Rabbit -
特異性
PDGFR alpha is membrane protein, so enrichment of membrane could help increasing the detection level of PDGFR alpha.
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アプリケーション
適用あり: WB, IHC-P, ICC/IF, Flow Cyt, IP, Indirect ELISA, IHC-Frmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: SH-SY5Y, A-204, MG-63, C6, and NIH/3T3 whole cell lysates. IHC-P: Mouse E14.5 lung, E14.5 intervertebral disc and uterus tissues; Rat E14.5 intervertebral disc tissue and Human endometrium tissue. ICC/IF: SH-SY5Y and A-204 cells. Flow Cyt: NIH/3T3 and A-204 cells. IP: A-204 whole cell lysate IHC-Fr: Human kidney tissue sections.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR22059-270 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab203491の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
1/1000. Detects a band of approximately 150, 180 kDa (predicted molecular weight: 122 kDa).
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IHC-P | (1) |
Use a concentration of 0.26 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/500.
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Flow Cyt |
1/50.
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IP |
1/30.
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Indirect ELISA |
Use a concentration of 1 µg/ml.
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IHC-Fr | (1) |
1/500.
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特記事項 |
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WB
1/1000. Detects a band of approximately 150, 180 kDa (predicted molecular weight: 122 kDa). |
IHC-P
Use a concentration of 0.26 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
Flow Cyt
1/50. |
IP
1/30. |
Indirect ELISA
Use a concentration of 1 µg/ml. |
IHC-Fr
1/500. |
ターゲット情報
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機能
Receptor that binds both PDGFA and PDGFB and has a tyrosine-protein kinase activity. -
組織特異性
Expressed in primary and metastatic colon tumors and in normal colon tissue. Tumors may express a different isoform to that found in normal tissue. -
関連疾患
Note=A chromosomal aberration involving PDGFRA is found in some cases of hypereosinophilic syndrome. Interstitial chromosomal deletion del(4)(q12q12) causes the fusion of FIP1L1 and PDGFRA (FIP1L1-PDGFRA). -
配列類似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 protein kinase domain. -
細胞内局在
Membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 5156 Human
- Entrez Gene: 18595 Mouse
- Entrez Gene: 25267 Rat
- Omim: 173490 Human
- SwissProt: P16234 Human
- SwissProt: P26618 Mouse
- SwissProt: P20786 Rat
- Unigene: 74615 Human
see all -
別名
- Alpha-type platelet-derived growth factor receptor antibody
- CD140 antigen-like family member A antibody
- CD140a antibody
see all
画像
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Lanes 1-8 : Anti-PDGFR alpha antibody [EPR22059-270] (ab203491) at 1/1000 dilution
Lanes 9-12 : Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution
Lanes 1 & 5 & 9 : SH-SY5Y
Lanes 2 & 6 & 10 : Human brain tissue lysate at 20 µg
Lanes 3 & 7 & 11 : Human heart tissue lysate at 20 µg
Lanes 4 & 8 & 12 : Human 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 why is the actual band size different from the predicted?
Exposure time: 80 secondsBlocking/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.
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Lanes 1-10 : Anti-PDGFR alpha antibody [EPR22059-270] (ab203491) at 1/1000 dilution
Lanes 11-15 : Anti-PDGFR alpha antibody [EPR5480] (ab134123) at 1/1000 dilution
Lanes 1 & 6 & 11 : Rat heart tissue lysate
Lanes 2 & 7 & 12 : Rat lung tissue lysate
Lanes 3 & 8 & 13 : Mouse brain tissue lysate
Lanes 4 & 9 & 14 : Mouse heart tissue lysate
Lanes 5 & 10 & 15 : Mouse lung tissue lysate
Lysates/proteins at 20 µg per lane.
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 why is the actual band size different from the predicted?
Exposure time: 80 secondsBlocking/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.
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All lanes : Anti-PDGFR alpha antibody [EPR22059-270] (ab203491) at 1/1000 dilution
Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : PDGFRA knockout SH-SY5Y cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 122 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab203491 observed at 150 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab203491 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 ab203491 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.
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Immunohistochemical analysis of paraffin-embedded human human bladder carcinoma labeling PDGFR alpha with ab203491 at 0.26 µg/mL followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in human bladder carcinoma was observed. The section was incubated with ab203491 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 solution 2) for 20 mins was used.
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Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling PDGFR alpha with ab203491 at 0.26 µg/mL followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in human endometrium was observed. The section was incubated with ab203491 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 solution 2) for 20 mins was used.
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ab203491 staining PDGFR alpha in wild-type SH-SY5Y cells (top panel) and PDGFRA knockout SH-SY5Y cells (ab275335) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab203491 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-204 (human muscle rhabdomyosarcoma cell line) cells and A-172 (human brain glioblastoma cell line) labeling PDGFR alpha with ab203491 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing membranous and cytoplasmic staining in A-204 cell line.
Negative control: A-172 (PMID:8425771.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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All lanes : Anti-PDGFR alpha antibody [EPR22059-270] (ab203491) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 122 kDa
Observed band size: 150,180 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time : Lane 1: 3 minutes; Lane 2: 59 seconds.
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Immunohistochemical analysis of paraffin-embedded mouse uterus tissue labeling PDGFR alpha with ab203491 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on interstitial cells of mouse uterus (PMID: 25788664). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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PDGFR alpha was immunoprecipitated from 0.35 mg of A-204 (human muscle rhabdomyosarcoma cell line) whole cell lysate with ab203491 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203491 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: A-204 whole cell lysate 10 µg (Input).
Lane 2: ab203491 IP in A-204 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203491 in A-204 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds.
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Flow cytometric analysis of NIH/3T3 (mouse embyro fibroblast cell line) cell line labeling PDGFR alpha with ab203491 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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Indirect ELISA using ab203491 at varying antibody concentrations (4000~0 ng /ml) and Human PDGF Receptor alpha antigen at 1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution dilution was used as a secondary antibody.
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IHC image of PDGFR alpha staining in a section of frozen normal human kidney performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab203491, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Immunohistochemical analysis of paraffin-embedded rat E14.5 intervertebral disc tissue labeling PDGFR alpha with ab203491 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in mesenchymal cells of rat E14.5 intervertebral disc (PMID: 9199674). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of paraffin-embedded mouse E14.5 lung tissue labeling PDGFR alpha with ab203491 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in the mesenchyme of mouse E14.5 lung (PMID: 8681381). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of paraffin-embedded mouse E14.5 intervertebral disc tissue labeling PDGFR alpha with ab203491 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in mesenchymal cells of mouse E14.5 intervertebral disc (PMID: 9199674). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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All lanes : Anti-PDGFR alpha antibody [EPR22059-270] (ab203491) at 1/1000 dilution
Lane 1 : A-204 (Human muscle rhabdomyosarcoma cell line) whole cell lysate
Lane 2 : MG-63 (human osteosarcoma fibroblast cell line) whole cell lysate
Lane 3 : A-172 (human brain glioblastoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 122 kDa
Observed band size: 150,180 kDa why is the actual band size different from the predicted?
Exposure time: 92 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Flow cytometric analysis of A-172 (human brain glioblastoma cell line, left) and A-204 (human muscle rhabdomyosarcoma cell line, right) cell lines labeling PDGFR alpha with ab203491 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (22)
ab203491 は 22 報の論文で使用されています。
- Sosa BR et al. A Subset of Osteosarcoma Bears Markers of CXCL12-Abundant Reticular Cells. JBMR Plus 6:e10596 (2022). PubMed: 35309866
- Chen L et al. Remodeling of dermal adipose tissue alleviates cutaneous toxicity induced by anti-EGFR therapy. Elife 11:N/A (2022). PubMed: 35324426
- Bo-Yin Z et al. Unlocking the Recovery Potential: JMJD3 Inhibition-Mediated SAPK/JNK Signaling Inactivation Supports Endogenous Oligodendrocyte-Lineage Commitment Post Mammalian Spinal Cord Injury. Neurochem Res 46:792-803 (2021). PubMed: 33428096
- Zhang J et al. miR-224 aggravates cancer-associated fibroblast-induced progression of non-small cell lung cancer by modulating a positive loop of the SIRT3/AMPK/mTOR/HIF-1a axis. Aging (Albany NY) 13:10431-10449 (2021). PubMed: 33819917
- Xia S et al. Delta-like 4 is required for pulmonary vascular arborization and alveolarization in the developing lung. JCI Insight 6:N/A (2021). PubMed: 33830085