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S1014-62A Mouse Anti-Smoothelin

Specifications
References
Clone Type
Monoclonal
Host
Mouse
Source
Chicken
Swiss Prot
P53814
Isotype
IgG1
Clone Number
4A83 (R4A)
Grade
Purified
Applications
IC IHC WB
Crossreactivity
Ca Ch Fe Hu Mk Po
Shipping Temp
Blue Ice
Storage Temp
-20°C
Notes
BSA Free

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.
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
Conjugates
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