Recombinant Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E249] to Caveolin-1 - Caveolae Marker
- Suitable for: IHC-P, WB, ICC
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Caveolin-1 antibody [E249] - Caveolae Marker
See all Caveolin-1 primary antibodies -
Description
Rabbit monoclonal [E249] to Caveolin-1 - Caveolae Marker -
Host species
Rabbit -
Specificity
This antibody should recognize both alpha and beta form of Caveolin-1. -
Tested applications
Suitable for: IHC-P, WB, ICCmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Caveolin-1 aa 150-250. The exact sequence is proprietary.
Database link: Q03135 -
Positive control
- WB: A431, A549 and HeLa cell lysates. ICC: A431 and HeLa cells. IHC-P: Human urinary bladder and lung tissues; Rat colon tissue; Mouse testis tissue.
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General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
E249 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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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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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab32577 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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IHC-P | 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use 0.01M Sodium Citrate Buffer, pH 6.0. | |
WB | 1/1000 - 1/10000. Predicted molecular weight: 20 kDa. | |
ICC | Use at an assay dependent concentration. |
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
- Entrez Gene: 25404 Rat
- Omim: 601047 Human
- SwissProt: Q03135 Human
- SwissProt: P49817 Mouse
- SwissProt: P41350 Rat
- Unigene: 74034 Human
see all -
Alternative names
- BSCL3 antibody
- CAV antibody
- CAV1 antibody
see all
Images
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All lanes : Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577) 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: 20 kDa
Observed band size: 20 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32577 observed at 20 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab32577 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. A431 wild-type and CAV1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab32577 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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ab32577 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 ab32577 at 1/200 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 [E249] - Caveolae Marker (ab32577)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling Caveolin-1 with Purified ab32577 at 1:500 dilution (2.1 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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All lanes : Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577) at 1/1000 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: 20 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32577 observed at 20 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab32577 was shown to react with Caveolin-1 in wild-type HeLa cells. 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. ab32577 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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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Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577) + A431 cell lysate
Predicted band size: 20 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?
Additional bands at: 22 kDa. We are unsure as to the identity of these extra bands. -
Immunocytochemistry/Immunofluorescence analysis of A431 cells labelling Caveolin-1 (green) with ab32577 at 1/200. Cells were fixed with 100% methanol. 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 (red), an Alexa Fluor® 488 conjugated mouse anti-tubulin antibody (1/200). Nuclei counterstained with DAPI (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse testis tissue sections labeling Caveolin-1 with purified ab32577 at 1:500 dilution (2.1 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human urinary bladder tissue, staining Caveolin-1 with ab32577 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human lung tissue, staining Caveolin-1 with ab32577 at 1/250 µg/ml.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)Image from Kaarteenaho R et al., BMC Dev Biol. 2010 Nov 16;10:113. Fig 6.; doi:10.1186/1471-213X-10-113; 16 November 2010, BMC Developmental Biology 2010, 10:113
Immunohistochemical analysis of developing Human lung tissue, staining Caveolin-1 with ab32577 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
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.
References (11)
ab32577 has been referenced in 11 publications.
- Peng W et al. LINC81507 act as a competing endogenous RNA of miR-199b-5p to facilitate NSCLC proliferation and metastasis via regulating the CAV1/STAT3 pathway. Cell Death Dis 10:533 (2019). PubMed: 31296840
- Marsboom G et al. Aberrant Caveolin-1-Mediated Smad Signaling and Proliferation Identified by Analysis of Adenine 474 Deletion Mutation (c.474delA) in Patient Fibroblasts: A New Perspective in the Mechanism of Pulmonary Hypertension. Mol Biol Cell N/A:N/A (2017). PubMed: 28298490
- Marsboom G et al. Aberrant caveolin-1-mediated Smad signaling and proliferation identified by analysis of adenine 474 deletion mutation (c.474delA) in patient fibroblasts: a new perspective on the mechanism of pulmonary hypertension. Mol Biol Cell 28:1177-1185 (2017). PubMed: 28468941
- Hu L et al. Emodin and rhein decrease levels of hypoxia-inducible factor-1a in human pancreatic cancer cells and attenuate cancer cachexia in athymic mice carrying these cells. Oncotarget 8:88008-88020 (2017). PubMed: 29152137
- Copeland CA et al. A disease-associated frameshift mutation in caveolin-1 disrupts caveolae formation and function through introduction of a de novo ER retention signal. Mol Biol Cell 28:3095-3111 (2017). PubMed: 28904206