Thermo Fisher Scientific CA IX Monoclonal Antibody (ZM41)

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:200

-

Product Specifications

Host/Isotype

Mouse / IgG2b, kappa

Class

Monoclonal

Type

Antibody

Clone

ZM41

Immunogen

Recombinant human CAIX protein

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

200 µg/mL

Storage conditions

2-8°C

Shipping conditions

Wet ice

Product Specific Information

A recommended positive control tissue for this product is Kidney, 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.

Carbonic anhydrases are a family of zinc containing metalloproteins that catalyze the reversible hydration of CO2. Among these, carbonic anhydrase IX (CA IX) is anchored to the cell membrane and is expressed in the human gastrointestinal tract, chiefly in the stomach and gall bladder. It is interesting to note that CA IX is overexpressed in epithelial malignancies of the uterus, cervix, lung, breast, and kidney; none of the associated normal tissues express this isozyme. Preliminary data suggest consistent immunoreactivity for anti-CA IX in clear cell renal cell carcinoma (RCC) with demonstrated sensitivity of 85% to 100%.Anti-CA IX, together with antibodies against PAX-2, Ksp-cadherin, and CD117, contributes to a robust panel that can be used to help make this distinction. Strong diffuse-to-multifocal immunostaining for anti-CA IX is observed in the large majority of urothelial carcinomas as opposed to the extremely weak and focal immunoreactivity seen in collecting duct carcinoma (CDC). Anti-CA IX can thus aid in distinguishing between urothelial carcinoma and CDC

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

Carbonic anhydrase (CA) is an enzyme that assists rapid interconversion of carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. It is abundant in all mammalian tissues. There are many genes that are inducible by hypoxia, via HIF-1 alpha. CA IX is one of the most inducible genes because of its stability and location within the membrane. Carbonic anhydrases have a widespread role in regulating pH in normal tissues, by regulating hydrogen ion (H+) flux. The pH is important in cell death under hypoxia, thus a blockade of CA IX results in increased cell death under hypoxia. Therefore, CA IX has become a reliable histochemical marker of hypoxia.

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