Thermo Fisher Scientific Calponin-1 Recombinant Rabbit Monoclonal Antibody (ZR297), RAbMono

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

Ready-to-use 150-200 µL

-

Product Specifications

Species Reactivity

Human

Host/Isotype

Rabbit / IgG

Expression System

CHO cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

ZR297

Immunogen

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Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Purification

Protein A

Storage buffer

tris with BSA, NP-40

Contains

<0.1% sodium azide

Storage conditions

4° C

Shipping conditions

Wet ice

Product Specific Information

This product is diluted and in a ready-to-use formulation.

A recommended positive control tissue for this product is Breast, however positive controls are not limited to this tissue type.

The primary antibody is intended for laboratory professional use in the detection of the corresponding protein in formalin-fixed, paraffin-embedded tissue stained in manual qualitative immunohistochemistry (IHC) testing. This antibody is intended to be used after the primary diagnosis of tumor has been made by conventional histopathology using non-immunological histochemical stains.

Multiple isoelectric variants of calponin have been identified, however only two molecular weight isoforms exist; a 34 kDa form and a 29 kDa form. Expression of the 29 kDa form, I-calponin, is primarily restricted to muscle of the urogenital tract, whereas the higher molecular weight variant has been demonstrated in vascular and visceral smooth muscle. In Western blotting, this MAb reacts with only the 34 kDa form of calponin in extracts of human aortic medial smooth muscle and is unreactive with fibroblast extracts of cultivated human foreskin. Calponin is a calmodulin, F-actin and tropomyosin binding protein, which is thought to be involved in the regulation of smooth muscle contraction. Calponin expression is restricted to smooth muscle cells and has been shown to be a marker of the differentiated (contractile) phenotype of developing smooth muscle.

Antibody is used with formalin-fixed and paraffin-embedded sections. Pretreatment of deparaffinized tissue with heat-induced epitope retrieval or enzymatic retrieval is recommended. In general, immunohistochemical (IHC) staining techniques allow for the visualization of antigens via the sequential application of a specific antibody to the antigen (primary antibody), a secondary antibody to the primary antibody (link antibody), an enzyme complex and a chromogenic substrate with interposed washing steps. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with a particular antigen.

A positive tissue control must be run with every staining procedure performed. This tissue may contain both positive and negative staining cells or tissue components and serve as both the positive and negative control tissue. External Positive control materials should be fresh autopsy/biopsy/surgical specimens fixed, processed and embedded as soon as possible in the same manner as the patient sample (s). Positive tissue controls are indicative of correctly prepared tissues and proper staining methods. The tissues used for the external positive control materials should be selected from the patient specimens with well-characterized low levels of the positive target activity that gives weak positive staining. The low level of positivity for external positive controls is designed to ensure detection of subtle changes in the primary antibody sensitivity from instability or problems with the staining methodology. A tissue with weak positive staining is more suitable for optimal quality control and for detecting minor levels of reagent degradation.

Internal or external negative control tissue may be used depending on the guidelines and policies that govern the organization to which the end user belongs to. The variety of cell types present in many tissue sections offers internal negative control sites, but this should be verified by the user. The components that do not stain should demonstrate the absence of specific staining, and provide an indication of non-specific background staining. If specific staining occurs in the negative tissue control sites, results with the patient specimens must be considered invalid.

Target Information

Calponin, a thin filament associated protein is implicated in the regulation and modulation of smooth muscle contraction. It is capable of binding to actin, calmodulin, troponin C and tropomyosin. Calponin is expressed in smooth muscle and tissues containing significant amounts of smooth muscle. Two isoforms of calponin exist whose molecular weights are 34kDa and 29kDa. Expression of the 29kDa form is primarily restricted to muscle of the urogenital tract. The expression of calponin has also been demonstrated in myoepithelial cells from benign and malignant breast lesions. It stains smooth muscle, myoepithelial cells, myofibroblasts, keratinocytes and nerve fibers. It identifies myoepithelial cells in breast lesions, and helps differentiate breast collagenous spherulosis (positive) from adenoid cystic carcinoma. Adenoid cystic carcinoma in salivary gland tumors is calponin positive.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.