Actin, Alpha-Smooth Muscle FITC conjugated

Description:
Fluorescein ~ Monoclonal Mouse Anti-Human Alpha-Smooth Muscle Actin

Application:
Immunofluorescence 10-20 micrograms/mL.
Flow cytometry 1-5 micrograms/106 cells (not tested in our lab.)
The optimal dilution should be determined by the end user.


Immunogen:
The N-terminal decapeptide of alpha-smooth muscle actin.

Cellular Localization:
Cytoplasmic

Species Reactivity:
Human, Baboon, bovine, mouse, rat, chicken, Rabbits. Others not tested.

Recommended Positive Control:
Bowel

Epitope:
Acetyl group and the four amino acids on the terminal end of the peptide chain of alpha-smoooth muscle actin.

Presentation:
IgG1 conjugated with Fluorescein Isothiocyanate (FITC) in 50 mM potassium phosphate, 150 mM Sodium Chloride, bovine serum albumin, carrier polysaccharides, and 0.05% Sodium Azide, pH 7.2.

Aliquoting Instructions:
Do not dilute the entire reconsituted solution at once. Withdraw aliquots as needed with a micropipette and keep concentrated stock at 4°C. Dilute according to the particular application being used. In general, the 0.05M borate pH 8.0 containing 0.15M sodium chloride, 0.02% sodium azide, is a good dilutent to use with most antibodies. GENERAL INFO: When diluting for immunohistochemistry, ELISA or western blot, make the dilution in Primary Antibody Diluting Buffer (Cat. No. M35). Avoid diluting the entire contents of the vial at once since the diluted solution may have reduced stability.

Staining Procedure:
Immunofluorescence on paraffin-embedded and frozen sections, as well as on cell smears. The antibody may be used for 2 hours at room temperature on formalin-fixed paraffin-embedded tissue sections. These are guidelines only; optimal conditions should be determined by the individual laboratory.

Specificity:
This antibody reacts with the alpha-smooth muscle isoform of actin. It does not react with actin from fibroblasts (beta- and gamma-cytoplasmic), striated muscle (alpha-sarcomeric) and myocardium (alpha-myocardial).

Storage:
Refrigerate in dark at 4°C. Do not freeze.

References:
1.    Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillesen D, Gabbiani G. A monoclonal antibody against alpha-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103: 2787-96, 1986.
2.    Skalli O, Schürch W, Seemayer T, et al. Myofibroblasts from diverse pathologic settings are heterogenous in their content of actin isoforms and intermediate filament proteins. Lab Invest 60: 275-85,1989.
3.    Skalli O, Pelte M-F, Peclet M-C, et al. Alpha-smooth muscle actin, a differentiation marker of smooth muscle cells, is present in microfilamentous bundles of pericytes. J Histochem Cytochem 37: 315-21, 1989.
4.    Rannov-Jessen L, Van Deurs B, Celis JE, Petersen OW. Smooth muscle differentiation in cultured human breast gland stromal cells. Lab Invest 63: 532-43, 1990.
5.    Hasegawa T, Hirose T, Kudo E, Abe J, Hizawa K. Cytoskeletal characteristics of myofibroblasts in benign and reactive fibroblastic lesions. Virochows Archiv (A) Pathol Anat 416:375-82, 1990.
6.    Schmitt-Gräff A, Krüger S, Bochard F, Gabbiani G, Denk H. Modulation of alpha smooth muscle actin and desmin expression in perisinusoidal cells of normal and diseased human livers. Am J Pathol 138:1233-42, 1991.