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Calponin

IHC of Calponin on an FFPE Leiomyoma Tissue

Description

Calponin is a 34 kDa polypeptide that interacts with actin, tropomyosin, and calmodulin. It is involved in smooth-muscle contraction mechanisms and is restricted exclusively to smooth-muscle tissue. Calponin is a calcium-binding protein. Calponin tonically inhibits the ATPase activity of myosin in smooth muscle. Phosphorylation of calponin by a protein kinase (which is dependent upon calcium binding to calmodulin) releases the calponin’s inhibition of the smooth-muscle ATPase.


Calponin has been found to be useful in differentiating benign sclerosing lesions of the breast from Carcinoma. Calponin positivity has also been noted in Malignant Myoepithelioma and Pleomorphic Adenoma of Salivary Gland origin, as well as in Angiomatoid Malignant Fibrous Histiocytoma.

Antibody Type
Mouse Monoclonal
Clone
CALP-A6
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-Calponin (CALP-A6) 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 5120
Prediluted
Ready-To-Use
3.0 ml
BSB 5121
Prediluted
Ready-To-Use
7.0 ml
BSB 5122
Prediluted
Ready-To-Use
15.0 ml
BSB 5123
Concentrated
1:50-1:200
0.1 ml
BSB 5124
Concentrated
1:50-1:200
0.5 ml
BSB 5125
Concentrated
1:50-1:200
1.0 ml
BSB 5126
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. NP, Wan BC, et al. Appl. Immunohistochem. 1997;5(3):141-151
  2. Nagao T, Sugano I, Ishida Y, et al. Cancer. 1998;Oct1:83(7):1292-9
  3. Savara AT, Gown AM, Zarbo RJ, Mod Pathol. 1997;Nov;10(11):1093-1100
  4. Fanburg-Smith JC, Meittinen M, Hum Pathol. 1999;Nov;30(11):1336-43

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