重组Anti-Caveolin-1抗体[EPR15554] - N-terminal (ab192869)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15554] to Caveolin-1 - N-terminal
- Suitable for: ICC, WB, IHC-P, Flow Cyt, IP
- Knockout validated
- Reacts with: Mouse, Human
概述
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产品名称
Anti-Caveolin-1抗体[EPR15554] - N-terminal
参阅全部 Caveolin-1 一抗 -
描述
兔单克隆抗体[EPR15554] to Caveolin-1 - N-terminal -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt MouseHumanICC HumanIHC-P MouseHumanIP HumanWB Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: PC-3, A549, A431 and HeLa cell lysates. IHC-P: Human liver and squamous cell carcinoma of cervix tissues; mouse lung tissue. ICC: A673 and HeLa cells. Flow Cyt: NIH3T3 and HeLa cells.
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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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR15554 -
同种型
IgG -
研究领域
相关产品
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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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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab192869 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Mouse
Human
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ICC |
Human
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IHC-P |
Mouse
Human
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IP |
Human
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WB |
Human
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应用 | Ab评论 | 说明 |
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ICC |
1/300.
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WB |
1/10000 - 1/50000. Detects a band of approximately 17, 20 kDa (predicted molecular weight: 17, 20 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt |
1/120.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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IP |
1/30.
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说明 |
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ICC
1/300. |
WB
1/10000 - 1/50000. Detects a band of approximately 17, 20 kDa (predicted molecular weight: 17, 20 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt
1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
IP
1/30. |
靶标
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功能
May act as a scaffolding protein within caveolar membranes. Interacts directly with G-protein alpha subunits and can functionally regulate their activity (By similarity). Involved in the costimulatory signal essential for T-cell receptor (TCR)-mediated T-cell activation. Its binding to DPP4 induces T-cell proliferation and NF-kappa-B activation in a T-cell receptor/CD3-dependent manner. Recruits CTNNB1 to caveolar membranes and may regulate CTNNB1-mediated signaling through the Wnt pathway. -
组织特异性
Expressed in muscle and lung, less so in liver, brain and kidney. -
疾病相关
Defects in CAV1 are the cause of congenital generalized lipodystrophy type 3 (CGL3) [MIM:612526]; also called Berardinelli-Seip congenital lipodystrophy type 3 (BSCL3). Congenital generalized lipodystrophies are autosomal recessive disorders characterized by a near absence of adipose tissue, extreme insulin resistance, hypertriglyceridemia, hepatic steatosis and early onset of diabetes. -
序列相似性
Belongs to the caveolin family. -
翻译后修饰
The initiator methionine for isoform Beta is removed during or just after translation. The new N-terminal amino acid is then N-acetylated. -
细胞定位
Golgi apparatus membrane. Cell membrane. Membrane > caveola. Membrane raft. Colocalized with DPP4 in membrane rafts. Potential hairpin-like structure in the membrane. Membrane protein of caveolae. - Information by UniProt
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数据库链接
- Entrez Gene: 857 Human
- Entrez Gene: 12389 Mouse
- Omim: 601047 Human
- SwissProt: Q03135 Human
- SwissProt: P49817 Mouse
- Unigene: 74034 Human
- Unigene: 28278 Mouse
- Unigene: 470537 Mouse
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别名
- BSCL3 antibody
- CAV antibody
- CAV1 antibody
see all
图片
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All lanes : Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869) at 1/1000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : CAV1 knockout A431 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 17, 20 kDa
Observed band size: 21-24 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab192869 observed at 21-24 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab192869 was shown to react with Caveolin-1 in wild-type A431 cells in western blot. Loss of signal was observed when CAV1 knockout sample was used. Wild-type and CAV1 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBS-T (0.1% Tween®) before incubation with ab192869 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] 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 hour at room temperature before imaging.
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ab192869 staining Caveolin-1 in wild-type HeLa cells (top panel) and CAV1 knockout HeLa cells (ab255371) (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 ab192869 at 1/500 dilution 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). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869)
Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix tissue labeling Caveolin-1 using ab192869 at 1/4000 dilution (0.15 μg/ml). A prediluted HRP Polymer for Rabbit IgG was used as the secondary. Counterstain: Hematoxylin. Negative control also shown.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow cytometry overlay histogram showing wild-type HeLa (green line) and CAV1 knockout HeLa cells (ab255371) stained with ab192869 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab192869) (1x106 in 100μl at 0.04 μg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type HeLa - black line CAV1 knockout HeLa - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869) at 1/10000 dilution
Lane 1 : A431 cell lysate
Lane 2 : A549 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : CAV1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 17, 20 kDaLanes 1 - 4: Merged signal (red and green). Green - ab192869 observed at 20 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab192869 was shown to react with Caveolin-1 in wild-type HeLa. Loss of signal was observed when knockout cell line ab255371 (knockout cell lysate ab263806) was used. Wild-type and Caveolin-1 knockout samples were subjected to SDS-PAGE. ab192869 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 dilution and 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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All lanes : Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869) at 1/10000 dilution
Lane 1 : PC-3 cell lysate
Lane 2 : A549 cell lysate
Lane 3 : A431 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 17, 20 kDa -
Immunoprecipitation analysis of A431 cell lysate labeling Caveolin-1 using ab192869 at 1/30 dilution (Lane 1). PBS negative control (Lane 2). Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869)
Immunohistochemical analysis of paraffin-embeddedMouse lung tissue labeling Caveolin-1 using ab192869 at 1/4000 dilution (0.15 μg/ml). A prediluted HRP Polymer for Rabbit IgG was used as the secondary. Counterstain: Hematoxylin. Negative control also shown.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Caveolin-1 using ab192869 at 1/4000 dilution (0.15 μg/ml). A prediluted HRP Polymer for Rabbit IgG was used as the secondary. Counterstain: Hematoxylin. Negative control also shown.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of A-673 cells labelling Caveolin-1 with ab192869 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889, Alexa Fluor® 594-conjugated anti-Tubulin [DM1A] at a dilution of 1/200. Nuclei counterstained with DAPI (blue).
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Flow cytometry analysis of NIH3T3 cells labeling Caveolin-1 using ab192869 at a 1/120 dilution (Red). A Goat anti rabbit IgG (FITC) at 1/150 dilution was used as secondary antibody. Cells were fixed with 2% paraformaldehyde. Cells without incubation with primary antibody and secondary antibody Blue. Rabbit monoclonal IgG was used as isotype control (Black).
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (7)
ab192869 被引用在 7 文献中.
- Streleckiene G et al. miR-20b and miR-451a Are Involved in Gastric Carcinogenesis through the PI3K/AKT/mTOR Signaling Pathway: Data from Gastric Cancer Patients, Cell Lines and Ins-Gas Mouse Model. Int J Mol Sci 21:N/A (2020). PubMed: 32013265
- Jäger M et al. Inducible Knockdown of Endothelial Protein Tyrosine Phosphatase-1B Promotes Neointima Formation in Obese Mice by Enhancing Endothelial Senescence. Antioxid Redox Signal 30:927-944 (2019). PubMed: 29390191
- Chen W et al. Clinical implications of hypoxia-inducible factor-1a and caveolin-1 overexpression in isocitrate dehydrogenase-wild type glioblastoma multiforme. Oncol Lett 17:2867-2873 (2019). PubMed: 30854062
- Bunaciu RP et al. Retinoic acid and 6-formylindolo(3,2-b)carbazole (FICZ) combination therapy reveals putative targets for enhancing response in non-APL AML. Leuk Lymphoma 60:1697-1708 (2019). PubMed: 30570341
- Camolotto SA et al. FoxA1 and FoxA2 drive gastric differentiation and suppress squamous identity in NKX2-1-negative lung cancer. Elife 7:N/A (2018). PubMed: 30475207
- Costa Verdera H et al. Cellular uptake of extracellular vesicles is mediated by clathrin-independent endocytosis and macropinocytosis. J Control Release 266:100-108 (2017). WB . PubMed: 28919558
- Chung KJ et al. A self-sustained loop of inflammation-driven inhibition of beige adipogenesis in obesity. Nat Immunol 18:654-664 (2017). PubMed: 28414311