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A recommended positive control tissue for this product is Lymph Node, 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.
Ki-67 antigen is a nuclear, non-histone protein that is present in all stages of the cell cycle except G0. This characteristic makes Ki-67 an excellent marker for proliferating cells and is commonly used as one of the prognostic factors in cancer studies. A correlation has been demonstrated between Ki-67 index and the histo-pathological grade of neoplasms. Assessment of Ki-67 expression in breast and neuroendocrine tumors shows a correlation between tumor proliferation and disease progression, thus making it possible to differentiate high-risk patients. Ki-67 expression may also prove to be important for distinguishing between malignant and benign peripheral nerve sheath tumors. Ki-67 labeling index has been shown to be a prognostic marker in a number of neoplasms including neuroendocrine tumor and breast carcinoma. In general, Ki-67 is a good marker of proliferating cell populations.
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.
Ki-67 is a nuclear protein that is expressed during various stages in the cell cycle, particularly during late G1, S, G2, and M phases. The protein has a forkhead associated domain (FHA) through which it associates with euchromatin at the perichromosomal layer, the centromeric heterochromatin, and the nucleolus. Ki-67 is shown to have a cell cycle dependent topographical distribution with perinucleolar expression at G1, expression in the nuclear matrix at G2, and expression on the chromosomes during M phase. Ki-67 is commonly used as a proliferation marker because it is not detected in G0 cells, but increases steadily from G1 through mitosis. Ki-67 antibodies are useful in establishing the cell growing fraction in neoplasms. In neoplastic tissues, the prognostic value is comparable to the tritiated thymidine-labelling index. The correlation between low Ki-67 index and histologically low-grade tumors is strong. Ki-67 is routinely used as a neuronal marker of cell cycling and proliferation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: Antigen identified by monoclonal antibody Ki-67; Antigen KI-67; Proliferation marker protein Ki-67; proliferation-related Ki-67 antigen; protein phosphatase 1, regulatory subunit 105; RP11-380J17.2
Gene Aliases: KIA; MIB-; MIB-1; MKI67; PPP1R105
UniProt ID: (Human) P46013
Entrez Gene ID: (Human) 4288
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