Immunofluorescence analysis of Phospho-Caveolin 2 pTyr19 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 Phospho-Caveolin 2 pTyr19 Rabbit Polyclonal Antibody (PA1060) 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 membranous localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to residues M(14) A D D A (pY) S H H S G C(25) of mouse CAV2.|
|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 *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||2 ug/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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA1-060 detects phospho-caveolin-2 Y19 from human and mouse samples.
PA1-060 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~21 kDa protein representing phospho-caveolin-2 Y19 from extract from COS-7 cells transiently transfected with the mouse caveolin-2 gene. Immunofluorescent staining of phospho-caveolin-2 Y19 in NIH 3T3 cells using PA1-060 results in a distinct punctate staining pattern at the cell periphery.
The PA1-060 immunogen is a synthetic phosphopeptide corresponding to residues M(14) A D D A (pY) S H H S G C(25) of mouse CAV2. This peptide (Cat. # PEP-180) 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 multigene 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 protein 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 tissue types.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Src-induced phosphorylation of caveolin-2 on tyrosine 19. Phospho-caveolin-2 (Tyr(P)19) is localized near focal adhesions, remains associated with lipid rafts/caveolae, but no longer forms a high molecular mass hetero-oligomer with caveolin-1.
PA1-060 was used in immunocytochemistry and western blot to investigate the phosphorylation of caveolin 2 by c-Src
|Lee H,Park DS,Wang XB,Scherer PE,Schwartz PE,Lisanti MP||The Journal of biological chemistry (277:34556)||2002|