Recombinant Anti-Caveolin-1 antibody [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/IF, Flow Cyt (Intra), WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Caveolin-1 antibody [EPR15554] - N-terminal
See all Caveolin-1 primary antibodies -
Description
Rabbit monoclonal [EPR15554] to Caveolin-1 - N-terminal -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PC-3, A549, A431 and HeLa cell lysates. IHC-P: Human liver and squamous cell carcinoma of cervix tissues; mouse lung tissue. ICC/IF: A673 and HeLa cells. Flow Cyt (intra): NIH3T3 and HeLa cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
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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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR15554 -
Isotype
IgG -
Research areas
Associated products
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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
Applications
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.
Application | Abreviews | Notes |
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ICC/IF |
1/300.
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Flow Cyt (Intra) |
1/120.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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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 | (2) |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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Notes |
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ICC/IF
1/300. |
Flow Cyt (Intra)
1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
IP
1/30. |
Target
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Function
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. -
Tissue specificity
Expressed in muscle and lung, less so in liver, brain and kidney. -
Involvement in disease
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. -
Sequence similarities
Belongs to the caveolin family. -
Post-translational
modificationsThe initiator methionine for isoform Beta is removed during or just after translation. The new N-terminal amino acid is then N-acetylated. -
Cellular localization
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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Database links
- 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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Alternative names
- BSCL3 antibody
- CAV antibody
- CAV1 antibody
see all
Images
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All lanes : Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869) at 1/1000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : CAV1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole 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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Immunocytochemistry/ Immunofluorescence - Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869)
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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Intracellular Intracellular 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 Fluorr® 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.
Performed under reducing conditions.
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 - Anti-Caveolin-1 antibody [EPR15554] - N-terminal (ab192869)
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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Intracellular 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).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (10)
ab192869 has been referenced in 10 publications.
- Cai B et al. A Pilot Application of an iTRAQ-Based Proteomics Screen Estimates the Effects of Cigarette Smokers' Serum on RPE Cells With AMD High-Risk Alleles. Transl Vis Sci Technol 11:15 (2022). PubMed: 35138344
- Yuan H et al. 5,2'-Dibromo-2,4',5'-trihydroxydiphenylmethanone Inhibits LPS-Induced Vascular Inflammation by Targeting the Cav1 Protein. Molecules 27:N/A (2022). PubMed: 35566232
- 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
- Liu Y et al. JMY expression by Sertoli cells contributes to mediating spermatogenesis in mice. FEBS J 287:5478-5497 (2020). PubMed: 32279424
- 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