重组Anti-CD146抗体[EPR3208] - BSA and Azide free (ab210072)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3208] to CD146 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CD146抗体[EPR3208] - BSA and Azide free
参阅全部 CD146 一抗 -
描述
兔单克隆抗体[EPR3208] to CD146 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, ICC/IF, 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
- ICC/IF: A375 cells. IHC-Fr: Mouse spleen tissue. IHC-P: Melanoma, breast carcinoma vessel, urinary bladder transitional carcinoma vessel, glioma vessel, normal tonsil and normal spleen tissue. WB: HeLa and HAP1 cell lysates. Flow Cyt (intra): A375 and HUVEC cells.
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常规说明
ab210072 is the carrier-free version of ab75769.
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.
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: 7.2
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
EPR3208 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-CD146 antibody [EPR3208] (ab196448)
- PE Anti-CD146 antibody [EPR3208] (ab209298)
- PerCP Anti-CD146 antibody [EPR3208] (ab216361)
- Biotin Anti-CD146 antibody [EPR3208] (ab217072)
- APC Anti-CD146 antibody [EPR3208] (ab303023)
- HRP Anti-CD146 antibody [EPR3208] (ab303024)
- Alexa Fluor® 647 Anti-CD146 antibody [EPR3208] (ab305213)
- Alexa Fluor® 555 Anti-CD146 antibody [EPR3208] (ab307390)
- Alexa Fluor® 594 Anti-CD146 antibody [EPR3208] (ab310672)
- Anti-CD146 antibody [EPR3208] (ab75769)
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab210072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) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 72 kDa. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
靶标
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功能
Plays a role in cell adhesion, and in cohesion of the endothelial monolayer at intercellular junctions in vascular tissue. Its expression may allow melanoma cells to interact with cellular elements of the vascular system, thereby enhancing hematogeneous tumor spread. Could be an adhesion molecule active in neural crest cells during embryonic development. Acts as surface receptor that triggers tyrosine phosphorylation of FYN and PTK2, and a transient increase in the intracellular calcium concentration. -
组织特异性
Detected in endothelial cells in vascular tissue throughout the body. May appear at the surface of neural crest cells during their embryonic migration. Appears to be limited to vascular smooth muscle in normal adult tissues. Associated with tumor progression and the development of metastasis in human malignant melanoma. Expressed most strongly on metastatic lesions and advanced primary tumors and is only rarely detected in benign melanocytic nevi and thin primary melanomas with a low probability of metastasis. -
序列相似性
Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
Contains 2 Ig-like V-type (immunoglobulin-like) domains. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 4162 Human
- Entrez Gene: 84004 Mouse
- Entrez Gene: 78967 Rat
- Omim: 155735 Human
- SwissProt: P43121 Human
- SwissProt: Q8R2Y2 Mouse
- SwissProt: Q9EPF2 Rat
- Unigene: 599039 Human
see all -
别名
- A32 antigen antibody
- CD 146 antibody
- CD146 antibody
see all
图片
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Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
Lane 2: CD146 knockout HAP1 whole cell lysate (40 µg)
Lane 3: A375 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab75769 observed at 120-72 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab75769 was shown to specifically react with CD146 in wild-type HAP1 cells as signal was lost in CD146 knockout cells. Wild-type and CD146 knockout samples were subjected to SDS-PAGE. ab75769 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/10000 dilution for 1 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
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Intracellular Flow Cytometry analysis of A375 (human malignant melanoma) cells labeling CD146 with unpurified ab75769 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
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Immunocytochemistry/Immunofluorescence analysis of A375 (human malignant melanoma) cells labelling CD146 with purified ab75769 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
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All lanes : Anti-CD146 antibody [EPR3208] (ab75769) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MCAM CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 120 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 containing PBS, BSA, glycerol, and sodium azide (ab75769).
Lanes 1- 2: Merged signal (red and green). Green - ab75769 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab75769 was shown to react with CD146 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab261790 (CRISPR/Cas9 edited cell lysate ab256985) lane below 120kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and MCAM CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab75769 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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This IHC data was generated using the same anti-CD146 antibody clone, EPR3208, in a different buffer formulation (cat# ab75769).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD146 with purified ab75769 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry experiments were used to compare symptomatic carotid plaques (SC) and asymptomatic carotid plaques (AsC)
Asymptomatic lesions presented higher CD146+ pericyte infiltration, p
ab75769 used at 1/200 dilution.
(After Figure 2 of Davaine et al)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Intracellular Flow Cytometry analysis of HUVEC cells labelling CD146 with purified ab75769 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled 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 (ab75769).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of melanoma tissue labelling CD146 with unpurified ab75769 at 1/250. A HRP/AP polymerized secondary antibody was used.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma vessels tissue labelling CD146 with unpurified ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of urinary bladder transitional carcinoma vessels tissue labelling CD146 with unpurified ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of glioma vessels tissue labelling CD146 with unpurified ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal tonsil tissue labelling CD146 with unpurified ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal spleen tissue labelling CD146 with unpurified ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (11)
ab210072 被引用在 11 文献中.
- Newton VL et al. Targeting apoptosis signalling kinase-1 (ASK-1) does not prevent the development of neuropathy in streptozotocin-induced diabetic mice. PLoS One 9:e107437 (2014). WB, IHC ; Mouse . PubMed: 25329046
- Ubil E et al. Mesenchymal-endothelial transition contributes to cardiac neovascularization. Nature 514:585-90 (2014). STED micro, Histology, Immunomicroscopy, IHC ; Mouse . PubMed: 25317562
- Davaine JM et al. Osteoprotegerin, pericytes and bone-like vascular calcification are associated with carotid plaque stability. PLoS One 9:e107642 (2014). IHC-P ; Human . PubMed: 25259713
- Gamblin AL et al. Bone tissue formation with human mesenchymal stem cells and biphasic calcium phosphate ceramics: The local implication of osteoclasts and macrophages. Biomaterials 35:9660-7 (2014). IHC-P ; Mouse . PubMed: 25176068
- Kilcoyne KR et al. Fetal programming of adult Leydig cell function by androgenic effects on stem/progenitor cells. Proc Natl Acad Sci U S A 111:E1924-32 (2014). IHC-P . PubMed: 24753613