Technical datasheet
Description
The monoclonal antibody 7C8 recognizes rat caveolin-1, a membrane protein of ~22 kDa. Caveolae are sphingomyelin/cholesterol-rich membrane domains first discovered as membrane invaginations on the surface of endothelial and epithelial cells. Caveolae are present in most cells, but are especially abundant in adipocytes. In addition to caveolins only two major protein components of caveolae were identified, namely the semicarbazide sensitive amine oxidase (SSAO) and the scavenger receptor CD36. Caveolin cycles between the plasma membrane and intracellular compartments via the endocytotic pathway. Caveolin is involved in the rapid intracellular transport of newly synthesized cholesterol from the ER directly to the caveolae. Caveolin plays an important role in multiple signaling pathways, molecular transport and cellular proliferation and differentiation. Caveolin binds to endothelial nitric oxide synthase leading to enzyme inhibition. Furthermore caveolin is a candidate tumor suppressor gene in many tumors. The specific functions of caveolin-1/caveolae are highly cell and context dependent.
The monoclonal antibody 7C8 recognizes caveolin-1α as well as caveolin-1β, which are present in many tissues, like aorta, heart, muscle, lung, adipose white, brown and epidydimal fat.
The monoclonal antibody 7C8 can be used to immuno-isolate caveolae.
Immunogen
GLUT4-containing vesicles immunoadsorbed from low density microsomes of rat adipocytes (Sprague Dawley) (Ref 4)
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Species
Mouse IgG2b
Application
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F
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FC
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FS
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IA
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IF1,3
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IP1
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P
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W1,2
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application notes
W: Reduced sample treatment and SDS-PAGE was used. The band sizes are ~22 kDa (Caveolin-1β) and ~25 kDa (caveolin-1α) (Ref.1).
Use
For Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Positive control
Adipocytes (3T3-L1 adipocytes)
Negative control
Cytoplasmic extracts of adipocytes
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Souto, R et al; Immunopurification and characterization of rat adipocyte caveolae suggest their dissociation from insulin signaling. J Biol Chem 2003, 278: 18321
2. Orrù, C et al; ACAT-1, Cav-1 and PrP expression in scrapie susceptible and resistant sheep. Cent Eur J Biol 2010, 5: 31
3. Salani, B et al; IGF-IR internalizes with caveolin-1 and PTRF/Cavin in Hacat cells. PLoS One 2010, 5: e14157
4. Thiodis, G et al; Immunological Analysis of GLUT4-enriched Vesicles:Identification of novel proteins regulated by insulin and diabetes. J of Biol Chem 1993, 268:11691
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Also available
References
1. Souto, R et al; Immunopurification and characterization of rat adipocyte caveolae suggest their dissociation from insulin signaling. J Biol Chem 2003, 278: 18321
2. Orrù, C et al; ACAT-1, Cav-1 and PrP expression in scrapie susceptible and resistant sheep. Cent Eur J Biol 2010, 5: 31
3. Salani, B et al; IGF-IR internalizes with caveolin-1 and PTRF/Cavin in Hacat cells. PLoS One 2010, 5: e14157
4. Thiodis, G et al; Immunological Analysis of GLUT4-enriched Vesicles:Identification of novel proteins regulated by insulin and diabetes. J of Biol Chem 1993, 268:11691