重组Anti-PDGFR alpha + PDGFR beta抗体[Y92] - C-terminal (ab32570)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y92] to PDGFR alpha + PDGFR beta - C-terminal
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP, ELISA
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PDGFR alpha + PDGFR beta抗体[Y92] - C-terminal
参阅全部 PDGFR alpha+PDGFR beta 一抗 -
描述
兔单克隆抗体[Y92] to PDGFR alpha + PDGFR beta - C-terminal -
宿主
Rabbit -
特异性
Expression levels of the target protein vary with sample type and some optimisation may be required. -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP, ELISAmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: N-GST tagged Human PDGF Receptor beta (aa557 to 1106) recombinant protein, N-GST tagged Human PDGF Receptor alpha (aa550 to 1089) recombinant protein, NIH/3T3 cell lysate. SH-SY5Y cell lysate. Rat brain and heart tissue lysate. Mouse brain tissue lysate. Human fetal brain tissue lysate. Human skeletal muscle tissue lysate. ICC/IF: NIH/3T3 cells. IHC-P: Human prostatic carcinoma, lung cancer, breast and spleen tissue. Flow Cyt (intra): NIH/3T3 cells. IP: NIH/3T3 cell lysate.
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常规说明
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y92 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- HRP Anti-PDGFR alpha + PDGFR beta antibody [Y92] (ab195515)
- Alexa Fluor® 488 Anti-PDGFR alpha + PDGFR beta antibody [Y92] (ab196376)
- Alexa Fluor® 594 Anti-PDGFR alpha + PDGFR beta antibody [Y92] (ab206872)
- Alexa Fluor® 405 Anti-PDGFR alpha + PDGFR beta antibody [Y92] (ab206873)
- Anti-PDGFR alpha + PDGFR beta antibody [Y92] - Low endotoxin, Azide free (ab215978)
- Anti-PDGFR alpha + PDGFR beta antibody [Y92] - BSA and Azide free (ab271835)
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Compatible Secondaries
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32570于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/20.
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WB | (4) |
1/5000 - 1/20000. Predicted molecular weight: 124 kDa.
For samples expressing low levels of PDGFR beta, the amount of lysate loaded may need to be increased to allow detection. |
IHC-P | (8) |
1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. Optimisation of the IHC protocol may be required depending on the sample used. |
ICC/IF | (8) |
1/100.
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IP |
1/20.
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ELISA |
Use at an assay dependent concentration.
Primary antibody concentration range: 1000 - 0 ng/mL |
说明 |
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Flow Cyt (Intra)
1/20. |
WB
1/5000 - 1/20000. Predicted molecular weight: 124 kDa. For samples expressing low levels of PDGFR beta, the amount of lysate loaded may need to be increased to allow detection. |
IHC-P
1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Optimisation of the IHC protocol may be required depending on the sample used. |
ICC/IF
1/100. |
IP
1/20. |
ELISA
Use at an assay dependent concentration. Primary antibody concentration range: 1000 - 0 ng/mL |
靶标
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细胞定位
PDGFR alpha: Membrane. PDGFR beta: Membrane. -
数据库链接
- Entrez Gene: 5156 Human
- Entrez Gene: 5159 Human
- Entrez Gene: 18595 Mouse
- Entrez Gene: 18596 Mouse
- Entrez Gene: 24629 Rat
- Entrez Gene: 25267 Rat
- Omim: 173410 Human
- Omim: 173490 Human
see all
图片
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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)
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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)
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All lanes : Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/1000 dilution
Lane 1 :Recombinant human PDGFR beta protein (ab60833)
Lane 2 :Recombinant human PDGFR alpha protein (ab84797)
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 124 kDa
Exposure time: 3 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution
Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : PDGFR beta knockout SH-SY5Y cell lysate
Lane 3 : Human Skeletal Muscle tissue lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 124 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?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.
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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.
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Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (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.
Immunofluorescence analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells stimulated with PDGF, staining PDGFR beta with unpurified ab32570.
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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.
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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.
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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.
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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.
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All lanes : Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/10000 dilution (purified)
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat heart tissue lysate
Lane 3 : Mouse brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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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.
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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.
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Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/10000 dilution (purified) + SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution (purified) + Human fetal brain tissue lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/50000 dilution (purified) + NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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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.
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (332)
ab32570 被引用在 332 文献中.
- Lin L et al. Mito-TEMPO, a Mitochondria-Targeted Antioxidant, Improves Cognitive Dysfunction due to Hypoglycemia: an Association with Reduced Pericyte Loss and Blood-Brain Barrier Leakage. Mol Neurobiol 60:672-686 (2023). PubMed: 36357613
- Şalva E et al. Combination therapy with chitosan/siRNA nanoplexes targeting PDGF-D and PDGFR-β reveals anticancer effect in breast cancer. J Gene Med 25:e3465 (2023). PubMed: 36413571
- Co CM et al. Biomolecule-releasing bioadhesive for glenoid labrum repair through induced host progenitor cell responses. J Orthop Res 41:1624-1636 (2023). PubMed: 36448179
- Liang X et al. Exosomal miR-532-5p induced by long-term exercise rescues blood-brain barrier function in 5XFAD mice via downregulation of EPHA4. Aging Cell 22:e13748 (2023). PubMed: 36494892
- Chen S et al. Three-dimensional mapping in multi-samples with large-scale imaging and multiplexed post staining. Commun Biol 6:148 (2023). PubMed: 36737476