重组Anti-PDGFR alpha + PDGFR beta抗体[Y92] - Low endotoxin,Azide free (ab215978)
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 - Low endotoxin, Azide free
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, ELISA, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-PDGFR alpha + PDGFR beta抗体[Y92] - Low endotoxin,Azide free -
描述
兔单克隆抗体[Y92] to PDGFR alpha + PDGFR beta - Low endotoxin,Azide free -
宿主
Rabbit -
特异性
Expression levels of the target protein vary with sample type and some optimisation may be required.
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经测试应用
适用于: ICC/IF, Flow Cyt (Intra), WB, ELISA, IP, IHC-Pmore 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
ab215978 is the carrier-free version of ab32570.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
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)
- PE Anti-PDGFR alpha + PDGFR beta antibody [Y92] (C-terminal) (ab307638)
- APC Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab314942)
- Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
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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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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab215978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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ICC/IF | (1) |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
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ELISA |
Use at an assay dependent concentration.
Primary antibody concentration range: 1000 - 0 ng/mL |
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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说明 |
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ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. |
ELISA
Use at an assay dependent concentration. Primary antibody concentration range: 1000 - 0 ng/mL |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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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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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 seconds
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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.
Observed band size: 170 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab32570).
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 loss of 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.
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ab32570 staining PDGFR alpha + 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
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ab32570 (purified) at 1/20 immunoprecipitating PDGFR alpha + 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
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ab32570 staining PDGFR alpha + 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
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Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling PDGFR alpha +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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
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Clone Y92 (ab215978) has been successfully conjugated by Abcam. This image was generated using Anti-PDGFR beta antibody [Y92] (Alexa Fluor® 488). Please refer to ab196376 for protocol details.
ab196376 staining PDGFR alpha + beta in NIH3T3 cells. The cells were fixed with 100% methanol (5 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab196376 at 1/50 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/167 dilution overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
Immunofluorescence analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells stimulated with PDGF, staining PDGFR alpha + beta with unpurified ab32570.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32570).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
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