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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)

  • Datasheet
  • SDS
Reviews (2)Q&A (1)References (12)

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Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunocytochemistry - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunocytochemistry - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
  • 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

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Overview

  • 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. This information is proprietary to Abcam and/or its suppliers.

  • 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.
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    E249
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • Coat Proteins
    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Caveolae and Clathrin
    • Cancer
    • Tumor biomarkers
    • Other
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab185043)
    • Alexa Fluor® 647 Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab193892)
    • HRP Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab193893)
    • Anti-Caveolin-1 antibody [E249] - BSA and Azide free (ab230262)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CAV1 (Caveolin-1) knockout HeLa cell line (ab255371)
    • Human CAV1 (Caveolin-1) knockout A-431 cell line (ab269583)
  • KO cell lysates

    • Human CAV1 (Caveolin-1) knockout HeLa cell lysate (ab263806)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human Caveolin-1 protein (ab114170)

Applications

The Abpromise guarantee

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
IHC-P (1)
1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use 0.01M Sodium Citrate Buffer, pH 6.0.
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 20 kDa.
ICC
Use at an assay dependent concentration.
Notes
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.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • 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
      modifications

      The 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.
    • Target information above from: UniProt accession Q03135 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • 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
      • Unigene: 28278 Mouse
      • Unigene: 470537 Mouse
      • Unigene: 22518 Rat
      see all
    • Alternative names

      • BSCL3 antibody
      • CAV antibody
      • CAV1 antibody
      • CAV1_HUMAN antibody
      • caveolae protein, 22 kD antibody
      • caveolin 1 alpha isoform antibody
      • caveolin 1 beta isoform antibody
      • Caveolin 1 caveolae protein 22kDa antibody
      • Caveolin-1 antibody
      • Caveolin1 antibody
      • cell growth-inhibiting protein 32 antibody
      • CGL3 antibody
      • LCCNS antibody
      • MSTP085 antibody
      • OTTHUMP00000025031 antibody
      • PPH3 antibody
      • VIP 21 antibody
      • VIP21 antibody
      see all

    Images

    • Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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 kDa



      Lanes 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.

    • Immunocytochemistry - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      Immunocytochemistry - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)

      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) - 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.

    • Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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 kDa



      Lanes 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.

       

    • Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      Western blot - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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 - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      Immunocytochemistry - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)

      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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      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.

    • Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
      Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)

    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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (12)

    Publishing research using ab32577? Please let us know so that we can cite the reference in this datasheet.

    ab32577 has been referenced in 12 publications.

    • Chen Y  et al. Irradiation pretreatment enhances the therapeutic efficacy of platelet-membrane-camouflaged antitumor nanoparticles. J Nanobiotechnology 18:101 (2020). PubMed: 32690018
    • 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
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Caveolin-1 antibody [E249] - Caveolae Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TRIS-EDTA-Buffer pH 9,0
    Permeabilization
    Yes - Wash Buffer from Dako with Tween
    Specification
    Kidney
    Fixative
    Paraformaldehyde
    Read More

    Mr. Rudolf Jung

    Verified customer

    Submitted Jun 17 2016

    Western blot abreview for Anti-Caveolin 1 antibody [E249]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HUVEC)
    Loading amount
    100000 cells
    Specification
    HUVEC
    Treatment
    various doses of VEGF for 24h
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 05 2008

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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