S1014-62A
Clone Type
MonoclonalHost
MouseSource
ChickenIsotype
IgG1Clone Number
4A83 (R4A)Grade
PurifiedApplications
IC IHC WBCrossreactivity
Ca Ch Fe Hu Mk PoShipping Temp
Blue IceStorage Temp
-20°CNotes
BSA Free
Mouse Anti-Smoothelin
Smoothelin is a constituent of the smooth muscle cell (SMC) cytoskeleton. Antibodies directed to smoothelin comprise a useful tool to monitor SMC differentiation. Smoothelin is exclusively expressed in fully differentiated (contractile) SMCs. RNA and protein analyses revealed a broad species distribution of this protein. Cells with SMC-like characteristics, such as myofibroblasts and myoepithelial cells as well as skeletal and cardiac muscle, do not contain smoothelin. Confocal scanning laser microscopy of tissue sections show a filamentous organization of smoothelin, colocalizing with actin stress fibers. In immunoblots two molecular weight isoforms are detected, i.e., a 59kD isoform specific for visceral SMC (Smoothelin A) and an isoform with a molecular weight of ~100kD in vascular SMC (Smoothelin B). Human smoothelin is encoded by a single copy gene which is located on chromosome 22. The monoclonal smoothelin antibody is useful for discriminating fully differentiated, contractile SMCs from proliferative SMCs. The distribution of smoothelin-positive cells indicates a correlation between smoothelin expression and smooth muscle tissue contractility. Smoothelin is a useful tool in the evaluation of atherosclerotic lesions.
Applications
Suitable for use in Western Blot, Immunocytochemistry and Immunohistochemistry. Other applications not tested.
Recommended Dilutions
Western Blot: 1:100-1:1000 Immunohistochemistry (Frozen, paraffin): 1:100-1:200, using ABC as detection reagent. For paraffin sections, pretreatment in 10mM citrate buffer, pH 6.0 and heating 3 times for 5 minutes in a microwave is required. Immunocytochemistry: permeabilized cells Optimum dilutions to be determined by researcher.
Hybridoma
Sp2/0-AG14 myeloma cells with spleen cells from Balb/c mice.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Cytoskeletal extract of chicken gizzard.
Form
Supplied as a liquid in PBS, pH 7.2, 0.09% sodium azide.
Purity
Purified from culture supernatant
Specificity
Recognizes chicken 59kD and 100kD protein, corresponding to smoothelin A and B, respectively, which are exclusively found in smooth muscle cells. Species Crossreactivity: human, canine, feline, porcine and monkey. Not with hamster or mouse.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
References
1. Van der Loop, F.T.L., et al., J. Cell Biol. 134: 401-411 (1996). 2. Wehrens, X.H.T., et al., FEBS Lett. 405: 315-320 (1997). 3. Van der Loop, F.T.L., et al., Arterioscler. Thromb. Vasc. Biol. 17: 665-671 (1997). 4. Christen, T., et al., Circ. Res. 85: 99-107 (1999). 5. Johansson, B., et al., Histochem. Cell Biol. 112: 291-299 (1999). 6. Johansson, B., et al., Histochem. J. 31: 723-727 (1999). 7. Deruiter, M.C., et al., Dev. Dyn. 221: 460-463 (2001). 8. Council, L., et al., (2009) Modern Pathology 22: 639-50. 9. Coco, D. P., et al., (2009). Am J Surg Pathol 33: 1795-1801.USBio References
No references available