Proliferation Marker Antibodies - FocusOn 056
Antibody tools for detection of proliferating cells
Antibodies are available for all markers listed overleaf. The highlight of the Acris Antibodies Ki-67 antibody panel is the rabbit monoclonal antibody clone SP6 (Figure 1 and Table 1). It is reactive in human and also in the rodent system and stains very strongly on paraffin sections.
In spite of the fact that PCNA is not the optimal marker for proliferation monitoring (see right-hand text) the PC10 antibody is helpful because it can be used on nearly all species, in contrast to the Ki-67 antibodies which are applicable in a subset of mammalian species only. For datasheet download and pricing info visit us at www.acris-antibodies.com.
Ki-67 is a 360 kDa protein and the prototypic cell cycle related nuclear protein, expressed by proliferating cells in all phases of the active cell cycle (G1, S, G2 and M phase). It is absent in resting (G0) cells. In G1 phase the Ki-67 antigen is predominantly localized in the perinucleolar region, in the later phases of the cell cycle the antigen is also detected throughout the nuclear interior, being mainly found in the nuclear matrix. In mitosis, the Ki-67 antigen is present on all chromosomes and appears in a reticulate structure surrounding the metaphase chromosomes. Because immunostaining rapidly decreases during anaphase and telophase, it has been concluded that Ki-67 is degraded with a biological half-life of the detectable antigen of less than one hour, as estimated in postmitotic cells during stathmokinesis induced by vinblastine. In contrast to many other cell cycle associated proteins like PCNA, the Ki-67 antigen is consistently absent in quiescent cells and is not detectable during DNA repair processes. Thus, the presence of the Ki-67 antigen is strictly associated with the cell cycle and confined to the nucleus, suggesting an important role of this structure in the maintenance and/or regulation of the cell division cycle.
Fig. 1: Staining of human tonsil paraffin sections using
Fig. 2: Staining of human tonsil paraffin sections using
Expression of the 36 kDa proliferating cell nuclear antigen (PCNA) is elevated in the nucleus during late G1 phase immediately before the onset of DNA synthesis, becoming maximal during S-phase and declining during G2 and M phases. Its level correlates directly with rates of cellular proliferation and DNA synthesis. PCNA may act as an auxiliary protein of DNA polymerase delta to play a fundamental role in the initiation of cell proliferation. A major drawback for PCNA as a proliferation marker for diagnostics is the fact that it is also detectable in almost all quiescent cells adjacent to some tumors and, presumably due to its long biological half-life, is also noticed in many quiescent tumor cells of different entities. A possible reason for the occurrence of PCNA in quiescent cells is its involvement, together with enzymes of the DNA replication machinery, in nucleotide excision repair mechanisms, as demonstrated in quiescent cells after ultra-violet irradiation in vitro and in vivo.
Fig. 3: Human tonsil (paraffin sections) stained with
Fig. 4: Western blot using affinity purified anti PCNA antibody (Cat.-No. AP09347PU-N) shows detection of PCNA protein in HEK293 (lane 1) and Jurkat (lane 2) whole cell extracts
The P105 protein, found in all human cells in a dimeric form (105 kDa) and monomeric form (41”¯kDa), is a proliferation-associated nuclear antigen that is absent in non-cycling cells in the G0 phase. This protein is important in the synthesis and transport of RNA in the cell cycle. During G2 and mitosis, this protein is increased possibly due to the unmasking of nucleolar antigens, resulting in large accumulations of this protein in the mitotic cytoplasm.
IPO-38 antigen is present in the nuclei of proliferating cells such as Hodgkin’s disease and non-Hodgkin’s lymphomas, different forms of leukemias, breast and colorectal carcinomas, and PHA-stimulated lymphocytes. It is not expressed in the cells of non-stimulated lymphocytes and granulocytes. IPO-38 antibodies can be a useful marking cell proliferation during monitoring of tumor progression.
Fig. 5: Human tonsil (paraffin sections) stained with
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|Rabbit||IgG||SP6||Supernatant||Bov, Hu, Ms, Rt||P||
1 ml / $610.00
|Acris Antibodies GmbH|
|+2 additional images||
Acris recommends this product for Immunohistochemistry.
|Mouse||IgG2a||PC10||FITC||Amph, Dros, Fi, Mam, Ye||F, ICC/IF||
0.1 mg / $400.00
|Acris Antibodies GmbH|
|Mouse||IgG2a||PC10||Purified||Hu, Insect, Ms, Rt, Ye||C, ICC/IF, P, WB||
1 ml / $380.00
|Acris Antibodies GmbH|
|+2 additional images|