Validated using a knockout cell line
Recombinant
RabMAb

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

Reviews (2)Q&A (1)References (10)

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: ICC/IF, IHC-P, ICC, WBmore 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/IF: A431 cells. IHC-P: Human urinary bladder and lung tissues; Rat colon tissue; Mouse testis tissue.

  • General notes

    A trial size is available to purchase for this antibody.

    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

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
ICC/IF Use at an assay dependent concentration. PubMed: 19808975
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use 0.01M Sodium Citrate Buffer, pH 6.0.
ICC 1/100.
WB 1/1000 - 1/10000. Predicted molecular weight: 20 kDa.
  • 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.
    • Information by UniProt

    • Database links

      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)

      Lane 1: A431 cell lysate (20 µg)
      Lane 2: A549 cell lysate (20 µg)
      Lane 3: Wild-type HeLa cell lysate (20 µg)
      Lane 4: CAV1 knockout HeLa cell lysate (20 µg)
      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. Loss of signal was observed when knockout sample 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.

    • 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)
      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.

    • 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.

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

      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.

    References

    This product has been referenced in:

    • 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). Read more (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 on the mechanism of pulmonary hypertension. Mol Biol Cell 28:1177-1185 (2017). Read more (PubMed: 28468941) »
    See all 10 Publications for this product

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

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

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

     Excellent
    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

    Question

    I have purchased several anti-bodies from abcam:
    ab83015
    ab79397
    ab81127
    ab48139
    ab32577
    ab98975.
    I have used the products WB but would now like to use them for IHC. The website says that they have not been tested in this application. Do you know how likely these Abs will be in succeeding if I attempt to do this
    Any help would be much appreaciated.

    Read More
    Answer

    Thank you for contacting us.

    Regarding antibodies ab79397 and ab32577, they both have been tested in IHC with paraffin embedded sections in the species stated on the datasheet. We don’t have preliminary data using them with frozen sections, but they are very likely to react in such conditions.

    Ab48139 has recently been validated to use in IHC-P in Canine, Feline, Mouse, Rat and Human.

    As for ab81127, ab83015 and ab98975, unfortunately, we have not characterised them by IHC, it doesn’t mean they won’t work in this application, but we don’t have data to support it.

    At Abcam we have an antibody evaluation system, called Abreview, by which customers interested in testing an antibody in new species or applications (not yet tested) can benefit from a free primary antibody in their next purchase if they share the results with us, publishing them in our website.

    To participate in this system, customers should first let us know the antibody they want to test in the new species/ applications. We then issue the discount code valid for one free primary antibody. This code must be issued before purchasing the antibody. The customer then purchases the antibody normally, and tests it in the new species / applications.

    Once we receive the results in our Website as an Abreview, the discount code becomes active, and it can be redeemed for any primary antibody ordered within the next 4 months.

    As mentioned, the discount code might be issued before the purchase of the antibody, but, as an exceptional case, I can still send a discount code if you would like to test any of these antibodies (ab81127, ab83015 and ab98975) with IHC and share your results with us.

    To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: http://it.adminsite.com/admin/crm2/www.abcam.com/collaborationdiscount.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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

     Excellent
    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

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