Figure 1. Caveolin-1 protein from human (HeLa), monkey (COS-7), Hamster (CHO), mouse (NIH/3T3), and rat (C6) cells was detected using PathScan® Total Caveolin-1 Sandwich ELISA Kit. However, this kit has a low detection level for Caveolin-1 in C2C12 cells and has no detection in PC-12 cells. The absorbance readings at 450 nm are shown in the top figure, while corresponding western blots using Caveolin-1 (D46G3) XP® Rabbit mAb #3267 are shown in the bottom figure.
Figure 2. The relationship between protein concentration of lysates from HeLa cells and the absorbance at 450 nm is shown. HeLa cells (85% confluence) were harvested and then lysed.
|REACTIVITY||H M R Hm Mk|
|Product Includes||Volume||Solution Color|
|Caveolin-1 Rabbit mAb Coated Microwells||96 tests|
|Caveolin-1 Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|TMB Substrate 7004||11 ml|
|STOP Solution 7002||11 ml|
|Sealing Tape||2 ea|
|ELISA Wash Buffer (20X) 9801||25 ml|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) 9803||15 ml|
The PathScan® Total Caveolin-1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Caveolin-1 protein. A Caveolin-1 Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Caveolin-1 protein is captured by the coated antibody. Following extensive washing, Caveolin-1 Mouse Detection mAb is added to detect the captured Caveolin-1 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of total Caveolin-1 protein.
Antibodies in kit are custom formulations specific to kit.
NOTE: Prepare solutions with purified water.
*NOTE: Some PathScan® ELISA Kits may include HRP-Linked Streptavidin in place of HRP-Linked Antibody.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
posted November 2013
Protocol Id: 204
PathScan® Total Caveolin-1 Sandwich ELISA Kit recognizes endogenous levels of Caveolin-1 protein as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.Species Reactivity:
Human, Mouse, Rat, Hamster, Monkey
The 21-24 kDa integral proteins, caveolins, are the principal structural components of the cholesterol/sphingolipid-enriched plasma membrane microdomain caveolae. Three members of the caveolin family (caveolin-1, -2, and -3) have been identified with different tissue distributions. Caveolins form hetero- and homo-oligomers that interact with cholesterol and other lipids (1). Caveolins are involved in diverse biological functions, including vesicular trafficking, cholesterol homeostasis, cell adhesion, and apoptosis, and are also implicated in neurodegenerative disease (2). Caveolins interact with multiple signaling molecules such as Gα subunit, tyrosine kinase receptors, PKCs, Src family tyrosine kinases, and eNOS (1,2). It is believed that caveolins serve as scaffolding proteins for the integration of signal transduction. Phosphorylation at Tyr14 is essential for caveolin association with SH2 or PTB domain-containing adaptor proteins such as GRB7 (3-5). Phosphorylation at Ser80 regulates caveolin binding to the ER membrane and entry into the secretory pathway (6).
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PathScan is a trademark of Cell Signaling Technology, Inc.