Technical Data
S1014-62A
Smoothelin
Description:
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 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.
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. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. 
TypeIsotypeCloneGrade
MabIgG14A83Purified
SizeStorageShippingSourceHost
100ug-20°CBlue IceChickenMouse
Concentration:
~1mg/ml
Immunogen:
Cytoskeletal extract of chicken gizzard.
Purity:
Purified
Form
Supplied as a liquid in PBS, pH 7.2, 0.09% sodium azide.
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
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
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.