Overview

  • Product name
  • Description
    Rabbit polyclonal to Caveolin-3
  • Host species
    Rabbit
  • Specificity
    This antibody does not detect caveolin-1 or -2.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IHC-Fr, Flow Cyt, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse Caveolin-3 aa 1-19.
    Sequence:

    MMTEEHTDLEARIIKDIHC


    (Peptide available as ab4930)

  • Positive control
    • WB: Rat heart, mouse heart, rat skeletal muscle, rat cardiac muscle and mouse muscle tissue lysates. ICC/IF: HeLa, A-375 and C2C11 cells. IHC-P: Mouse heart, lymph node and skeletal muscle tissue sections. IP: Mouse heart tissue lysate. Flow Cyt: U-87 MG cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab2912 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 1/100 - 1/200.
ICC/IF 1/20.
IHC-Fr Use at an assay dependent concentration. PubMed: 21408028
Flow Cyt Use 3-5µg for 106 cells.
WB Use a concentration of 1 - 3 µg/ml. Can be blocked with Mouse Caveolin-3 peptide (ab4930).
IP Use at an assay dependent concentration.

Target

  • Function
    May act as a scaffolding protein within caveolar membranes. Interacts directly with G-protein alpha subunits and can functionally regulate their activity. May also regulate voltage-gated potassium channels. Plays a role in the sarcolemma repair mechanism of both skeletal muscle and cardiomyocytes that permits rapid resealing of membranes disrupted by mechanical stress.
  • Tissue specificity
    Expressed predominantly in muscle.
  • Involvement in disease
    Defects in CAV3 are the cause of limb-girdle muscular dystrophy type 1C (LGMD1C) [MIM:607801]. LGMD1C is a myopathy characterized by calf hypertrophy and mild to moderate proximal muscle weakness. LGMD1C inheritance can be autosomal dominant or recessive.
    Defects in CAV3 are a cause of hyperCKmia (HYPCK) [MIM:123320]. It is a disease characterized by persistent elevated levels of serum creatine kinase without muscle weakness.
    Defects in CAV3 are a cause of rippling muscle disease (RMD) [MIM:606072]. RMD is a rare disorder characterized by mechanically triggered contractions of skeletal muscle. In RMD, mechanical stimulation leads to electrically silent muscle contractions that spread to neighboring fibers that cause visible ripples to move over the muscle.
    Defects in CAV3 are a cause of cardiomyopathy familial hypertrophic (CMH) [MIM:192600]; also designated FHC or HCM. Familial hypertrophic cardiomyopathy is a hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death.
    Defects in CAV3 are the cause of long QT syndrome type 9 (LQT9) [MIM:611818]. Long QT syndromes are heart disorders characterized by a prolonged QT interval on the ECG and polymorphic ventricular arrhythmias. They cause syncope and sudden death in response to excercise or emotional stress. They can present with a sentinel event of sudden cardiac death in infancy.
    Defects in CAV3 can be a cause of sudden infant death syndrome (SIDS) [MIM:272120]. SIDS is the sudden death of an infant younger than 1 year that remains unexplained after a thorough case investigation, including performance of a complete autopsy, examination of the death scene, and review of clinical history. Pathophysiologic mechanisms for SIDS may include respiratory dysfunction, cardiac dysrhythmias, cardiorespiratory instability, and inborn errors of metabolism, but definitive pathogenic mechanisms precipitating an infant sudden death remain elusive. Long QT syndromes-associated mutations can be responsible for some SIDS cases.
  • Sequence similarities
    Belongs to the caveolin family.
  • Cellular localization
    Golgi apparatus membrane. Cell membrane. Membrane > caveola. Potential hairpin-like structure in the membrane. Membrane protein of caveolae.
  • Information by UniProt
  • Database links
  • Alternative names
    • CAV3 antibody
    • CAV3_HUMAN antibody
    • Caveolin 3 antibody
    • Caveolin-3 antibody
    • LGMD1C antibody
    • LQT9 antibody
    • M-caveolin antibody
    • MGC126100 antibody
    • MGC126101 antibody
    • MGC126129 antibody
    • OTTHUMP00000115603 antibody
    • OTTHUMP00000207105 antibody
    • VIP 21 antibody
    • VIP21 antibody
    see all

Images

  • All lanes : Anti-Caveolin-3 antibody (ab2912)

    Lane 1 : Rat heart tissue lysate
    Lane 2 : Mouse heart tissue lysate
    Lane 3 : HEK293 cell lysate
    Lane 4 : Rat skeletal muscle tissue lysate
    Lane 5 : Mouse muscle tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG (H+L) at 1/2500 dilution

    Developed using the ECL technique.

    Observed band size: 17 kDa
    why is the actual band size different from the predicted?



    Blocked with 5% skimmed milk.

  • Immunocytochemistry/Immunofluorescence analysis of Caveolin-3 in HeLa Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) (right panel) or with ab2912 at a dilution of 1/20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Caveolin-3 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized mouse heart tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing Caveolin-3 ab2912 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Caveolin-3 was immunoprecipitated using 5 µg of ab2912 from mouse heart tissue lysate (Lane 3) using the protein A beads. Normal rabbit IgG was used as a isotype control (Lane 2). 10% input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using ab2912 and HRP-conjugated goat anti-rabbit IgG (H+L) at a dilution of 1/2500. Chemiluminescent detection was performed.

  • Flow cytometry analysis of U-87 MG cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with ab2912 (red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488-conjugated goat anti-rabbit secondary antibody at a dilution of 1/400 for 30 minutes at room temperature. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

  • Immunofluorescent analysis of Caveolin-3 in C2C11 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) (right panel) or with ab2912 at a dilution of 1/20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Caveolin-3 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse skeletal muscle tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/100 with a rabbit polyclonal antibody recognizing Caveolin-3 ab2912 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Anti-Caveolin-3 antibody (ab2912) + Rat cardiac muscle tissue lysate
  • Immunohistochemistry was performed on normal biopsies of deparaffinized mouse lymph node tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing Caveolin-3 ab2912 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunocytochemistry/Immunofluorescence analysis of 70% confluent log phase A-375 cells. Cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. Samples were incubated with ab2912 at 1µg/ml in 1% BSA for 3 hours at room temperature and then labelled with Alexa Fluor® 488-conjugated goat anti-rabbit IgG (H+L) at a dilution of 1/2000 for 45 minutes at room temperature (panel a: green). Nuclei (panel b: blue) were stained with DAPI. F-actin (panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (1/300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

References

This product has been referenced in:
  • Shen X  et al. 3D dSTORM imaging reveals novel detail of ryanodine receptor localization in rat cardiac myocytes. J Physiol 597:399-418 (2019). Read more (PubMed: 30412283) »
  • González Coraspe JA  et al. Biochemical and pathological changes result from mutated Caveolin-3 in muscle. Skelet Muscle 8:28 (2018). Read more (PubMed: 30153853) »
See all 39 Publications for this product

Customer reviews and Q&As

11-15 of 15 Abreviews or Q&A

Answer

I have looked into it and yes, we have had a mistake on the datasheet all along regarding the immunogen. It is based on Rat(Ms ) calveolin 3 sequence. I have updated the datasheet. This explains as why the specificity mentions rat tissues. This does not mean that the antibody is specific for rat tissues only, it was just one of the inital species tested. I have taken this off as it confuses people. During the years we have had the antibody, we have not had any customers contacting us regarding problems with Human samples. Also antibody has been consistently over the years a good seller. As mentioned before, we guarantee Human cross-reactivity. Moreover, we have references and Abreviews for human samples by other customers. I can see that you have contacted us earlier this year regarding background in overexpressed calveolin-3 WB. I would encourage you to send us the filled in WB questionnaire with images as we might be able to help you with protocol tips to improve your result.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (kidney,HEK)
Loading amount
15 µg
Specification
kidney,HEK
Treatment
transfected with WT-Cav3
Blocking step
Blocking buffer Roche as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2%

Abcam user community

Verified customer

Submitted Mar 05 2007

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Heart)
Specification
Heart
Fixative
Formaldehyde
Antigen retrieval step
None
Blocking step
Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 10

Abcam user community

Verified customer

Submitted Jun 29 2006

Answer

Thank you for your email. Since unfortunately none of the Caveolin 1 antibodies have been tested yet for cross-reactivity with rabbit, we would not be able to guarantee results and thus a refund would not apply. Should you decide to go ahead and purchase one of these products, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 points will be offered for an image). Please let me know if you require any further assistance.

Read More

Answer

Thank you for your enquiry. At the moment we have one Caveolin 3 antibody: ab2912 - Rabbit polyclonal to Caveolin 3 - Caveolae Marker. We have three Caveolin 1 antibodies: ab18199 - Rabbit polyclonal to Caveolin 1 - Caveolae Marker ab2910 - Rabbit polyclonal to Caveolin 1 - Caveolae Marker ab17052 - Mouse monoclonal [7C8] to Caveolin 1 None of these antibodies have been tested yet for cross-reactivity with rabbit or guinea pig. Please contact us again if you have any additional questions.

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11-15 of 15 Abreviews or Q&A

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