Analysis Note

ControlPOSITIVE CONTROL:

Smooth muscle (e.g. aterial tunica media). Negative control: any nonmuscle tissue (e.g. arterial tunica adentitial).

Application

Western blot. Suggested blocking buffer is TBS-Tween with 2% BSA. Suggested dilution buffer is TBS-Tween with 0.05% sodium azide. Preferred gel percentage is 7% and/or 4-20% (or similar) gradient gel.

Immunohistochemistry on frozen and paraffin embedded tissue sections. Suggested fixation for frozen tissue sections is acetone fix for 6 minutes at room temperature. For formalin fixed paraffin embedded tissue sections: microwave in 0.01M citrate buffer (pH 6.0) for 8-10 minutes (note that all microwaves differ and adjustments may need to be made) follow with enzyme digestion (0.01% pronase for 10 mintues). Suggested blocking agent is fetal bovine serum. The antibody has also been used successfully on methyl-Carnoy fixed tissue.

Immunocytochemistry

Immunoprecipitation. Suggested extraction buffer is 20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholic acid-NaCl and 0.5 mM PMSF. Final reaction volume is 1 mL and suggested capture agent is agarose conjugated anti-mouse IgG.

Optimal working dilutions must be determined by the end user.

Research Sub CategoryMuscle Physiology

Detect Caldesmon using this Anti-Caldesmon Antibody, smooth muscle, clone N5/22 validated for use in IP, WB, IC, IH(P).

Research CategoryMetabolism

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Caldesmon is a protein component of the thin filaments of smooth muscle myofibrils. It

is also localized in the stress fibers of fibroblasts. Caldesmon was identified as a

Ca2+/Calmodulin-binding protein with molecular weight of 120-150kDa (H-Caldesmon)

and 70-80kDa (L-Caldesmon). H-Caldesmon is the primary isoform in smooth muscle

while L-Caldesmon is most abundant in non-muscle cells. Caldesmon is capable of

binding two calmodulin molecules, one at either end of the protein. Additionally,

Caldesmon is an actin, myosin, and tropomyosin-binding protein. Human L-Caldesmon

is a protein of 538 amino acids with mobility of 80kDa. In vitro, L-Caldesmon inhibits

the actomyosin ATPase in an F-Actin-dependent manner. L-Caldesmon may play an

important function in motile processes such as secretion and organelle movement.

Immunogen

Epitope: smooth muscle

Crude smooth muscle extract from normal human adult uterus.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-590

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Liquid in 0.02M Phosphate buffer with 0.25M NaCl and 0.1% sodium azide.

Format: Purified

Specificity

Caldesmon, smooth muscle. By Western blot the antibody recognizes a protein of 150-kDa.

Storage and Stability

Maintain at 2-8°C in undiluted aliquots up to 6 months after date of receipt