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Caldesmon

IHC of Caldesmon on an FFPE Appendix Tissue

Description

Caldesmon 1, also known as CALD1, is a human gene. Caldesmon is a calmodulin-binding protein. Like Calponin, Caldesmon tonically inhibits the ATPase activity of myosin in smooth muscle. This gene encodes a Calmodulin and actin-binding protein that play an essential role in the regulation of smooth muscle and nonmuscle contraction.


Two closely-related variants of human Caldesmon have been identified. The h-Caldesmon variant (120–150 kD) is predominantly expressed in smooth muscle, whereas I-Caldesmon (70–80 kD) is found in non-muscle tissue and cells. Neither of the two variants has been detected in skeletal muscle. Anti-Caldesmon recognizes only the h-Caldesmon variant. Anti-Caldesmon antibody labels smooth muscle and tumors of smooth muscle, myofibroblastic, and myoepithelial differentiation. Anti-Caldesmon has also been used to differentiate Epithelioid Mesothelioma from Serous Papillary Carcinoma of the ovary.

Antibody Type
Mouse Monoclonal
Clone
CALD-31
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Appendix, Uterus, Leiomyoma
Storage
Store at 2°-8°C
Stability
2 years

For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).

Presentation

Anti-Caldesmon (CALD-31) is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 6099
Prediluted
Ready-To-Use
3.0 ml
BSB 6100
Prediluted
Ready-To-Use
7.0 ml
BSB 6101
Prediluted
Ready-To-Use
15.0 ml
BSB 6102
Concentrated
1:100-1:400
0.1 ml
BSB 6103
Concentrated
1:100-1:400
0.5 ml
BSB 6104
Concentrated
1:100-1:400
1.0 ml
BSB 6105
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Huber PA, Int J. Biochem Cell Biol. 1998;29(8-9):1047-51
  2. Hisaoka M, et al. Appl. Immunohistochem. Mol. Morphol. 2003;9(4):302-8
  3. Frid MG, et al. Dev Biol. 1992;153:185
  4. Sobue K, Sellers JR, J Biol Chem. 1991;266(19):12115
  5. Comin CE, et al. Am J Surg Pathol. 2007;Aug;3198):1139-48

Recommended Immunohistochemical Protocol

Pretreatment
  1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
  2. Air dry for 2 hours at 58° C.
  3. Deparaffinize, dehydrate and rehydrate tissues.
  4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
  5. Any of three heating methods may be used:
    • Electric Pressure Cooker
      Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
    • Water Bath Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
    • Conventional Steamer Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
  6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
  7. Wash slides with IHC wash buffer or DI water.
  8. Continue IHC staining protocol.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain