Anti-Caldesmon Antibody, smooth muscle, clone N5/22

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.

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

Crude smooth muscle extract from normal human adult uterus.

Epitope: smooth muscle

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

Research Category
Metabolism

Research Sub Category
Muscle Physiology

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.

Replaces: 04-590

Format: Purified

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

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

Control
POSITIVE CONTROL:

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

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

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