Immunofluorescence analysis of Caveolin 3 was done on 70% confluent log phase A-375 cells. The 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. The cells were labeled with Caveolin 3 Rabbit Polyclonal Antibody (PA1066) at 1ug/ml in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 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
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) M T E E H T D L E A R I I K D I H C(19) C of mouse CAV3|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunoprecipitation (IP)||5 µg|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-066 detects Caveolin 3 from rat and mouse tissues. This antibody does not detect caveolin-1 or -2.
PA1-066 has been successfully used in Western blot, IF, immunocytochemistry and immunoprecipitation procedures. By Western blot, this antibody detects an ~21 kDa protein representing Caveolin 3 from rat heart homogenate.
The PA1-066 immunogen is a synthetic peptide corresponding to residues M(1) M T E E H T D L E A R I I K D I H C(19) C of mouse CAV3. This sequence is ~85% conserved in human Caveolin 3. PA1-066 immunizing peptide (Cat. # PEP-081) is available for use in neutralization and control experiments.
Caveolae are specialized domains of the plasma membrane that are implicated in the sequestration of a variety of lipid and protein molecules. It has been suggested that these important cellular organelles have a pivotal role in such diverse biochemical processes as lipid metabolism, growth regulation, signal transduction, and apoptosis. Caveolin interacts with and regulates heterotrimeric G-proteins. Currently, there are three members of the caveolin gene family which are known to encode 21-24 kDa integral membrane proteins that comprise the major structural component of the caveolar membrane in vivo. Caveolin-2 is abundantly expressed in fibroblasts and differentiated adipocytes, smooth and skeletal muscle, and endothelial cells. The expression of caveolin-1 is similar to that of caveolin-2 while caveolin-3 expression appears to be limited to muscle tissues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Ca2+ sparks and cellular distribution of ryanodine receptors in developing cardiomyocytes from rat.
PA1-066 was used in immunocytochemistry to examine the role of ryanodine receptors in the development of rat cardiomyocytes
|Snopko RM,Ramos-Franco J,Di Maio A,Karko KL,Manley C,Piedras-Rentería E,Mejía-Alvarez R||Journal of molecular and cellular cardiology (44:1032)||2008|
Loss of caveolin-3 induced by the dystrophy-associated P104L mutation impairs L-type calcium channel function in mouse skeletal muscle cells.
PA1-066 was used in immunocytochemistry to study the role of caveolin-3 in L-type calcium channel membrane function and localization in skeletal muscle cells .
|Couchoux H,Allard B,Legrand C,Jacquemond V,Berthier C||The Journal of physiology (580:745)||2007|
Role of caveolae in signal-transducing function of cardiac Na+/K+-ATPase.
PA1-066 was used in immunocytochemistry to study the function of cardiac sodium/potassium-ATPase
|Liu L,Mohammadi K,Aynafshar B,Wang H,Li D,Liu J,Ivanov AV,Xie Z,Askari A||American journal of physiology. Cell physiology (284:C1550)||2003|
Phosphofructokinase muscle-specific isoform requires caveolin-3 expression for plasma membrane recruitment and caveolar targeting: implications for the pathogenesis of caveolin-related muscle diseases.
PA1-066 was used in western blot to study the role of muscle caveolae in the control of energy metabolism in skeletal muscle fibers.
|Sotgia F,Bonuccelli G,Minetti C,Woodman SE,Capozza F,Kemp RG,Scherer PE,Lisanti MP||The American journal of pathology (163:2619)||2003|
Intracellular retention of glycosylphosphatidyl inositol-linked proteins in caveolin-deficient cells.
PA1-066 was used in western blot to investigate the effect of caveolin on glycosylphosphatidyl inositol-linked protein function .
|Sotgia F,Razani B,Bonuccelli G,Schubert W,Battista M,Lee H,Capozza F,Schubert AL,Minetti C,Buckley JT,Lisanti MP||Molecular and cellular biology (22:3905)||2002|
Synapse-associated protein 97 selectively associates with a subset of AMPA receptors early in their biosynthetic pathway.
PA1-066 was used in immunoprecipitation to investigate the interaction between GluR1 and SAP97.
|Sans N,Racca C,Petralia RS,Wang YX,McCallum J,Wenthold RJ||The Journal of neuroscience : the official journal of the Society for Neuroscience (21:7506)||2001|