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
Crude smooth muscle extract from normal human adult uterus.
Epitope: smooth muscle
Application
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.
Biochem/physiol Actions
Caldesmon, smooth muscle. By Western blot the antibody recognizes a protein of 150-kDa.
Physical form
Format: Purified
Liquid in 0.02M Phosphate buffer with 0.25M NaCl and 0.1% sodium azide.
Preparation Note
Maintain at 2-8°C in undiluted aliquots up to 6 months after date of receipt
Analysis Note
Control
POSITIVE CONTROL:
Smooth muscle (e.g. aterial tunica media). Negative control: any nonmuscle tissue (e.g. arterial tunica adentitial).
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Replaces: 04-590
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
| biological source | mouse |
| Quality Segment | 100 |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | N5/22, monoclonal |
| species reactivity | rabbit, bovine, human, pig |
| manufacturer/tradename | Chemicon® |
| technique(s) | immunocytochemistry: suitable,immunohistochemistry: suitable (paraffin),immunoprecipitation (IP): suitable,western blot: suitable |
| isotype | IgG1 |
| NCBI accession no. | NM_004342.5,NM_033138.2,NM_033139.2,NM_033140.2,NM_033157.2 |
| UniProt accession no. | Q05682 |
| shipped in | wet ice |
| target post-translational modification | unmodified |
| Gene Information | human ... CALD1(800) |
