Ki67 is a nuclear protein expressed throughout the energetic phases of the cell cycle, which makes it a biomarker of cell proliferation. In medical pathology settings, immunohistochemical (IHC) detection of Ki67 is used to calculate Ki67 proliferation indices (PIs), which have prognostic info and are used to subdivide breast carcinomas and neuroendocrine neoplasias. Calculation of Ki67 PIs is notoriously arduous and vulnerable to intraobserver and interobserver variance.
As well as, IHC protocol settings [such as primary antibody (Ab) clone, clone format, and stainer platform] can have an effect on the results of the IHC assays and in flip the Ki67 PI. Digital picture evaluation has been prompt as a great tool to standardize Ki67 counting. Just lately, digital double staining, a pc algorithm segmenting Ki67 and Ki67 tumor cells utilizing digitally fused parallel cytokeratin and Ki67-stained slides, has been launched.
On this examine, we evaluate Ki67 PIs obtained by digital double staining in 41 breast carcinomas stained utilizing probably the most generally used commercially accessible major Ab clones and codecs on the principle stainer platforms. IHC protocols for the concentrated (conc) Ab and platform mixtures had been optimized for the very best analytical sensitivity and optimum signal-to-noise ratio, whereas ready-to-use (RTU) codecs had been used, as beneficial by the seller.
Vital variations within the imply Ki67 PIs (relativized to the imply core Ki67) had been noticed not solely between the totally different Ab clones but in addition the totally different codecs and stainer platforms; Ki67 PIs with SP6 conc stained on the Ventana BenchMark ULTRA platform had been on common 11.9 proportion factors (pp) larger than the imply core common, whereas with Ab 30.9 RTU on the Ventana platform, they had been 10.
Four pp larger. Mib1 RTU (Dako Autostainer Hyperlink 48) and MM1 RTU (Leica Bond) supplied 8.6 and 12.5 pp decrease Ki67 PIs, respectively. Mib1 conc and SP6 conc on the Dako Autostainer and Leica Bond supplied comparable results-close to the general common. Vital variations within the proportion of tumors with Ki67 high-level expression (Ki67 PI ≥20%) had been noticed amongst Ab, format, and stainer platform mixtures.
The outcomes underline the challenges within the comparability of Ki67 PIs throughout Abs, codecs, and platforms. Researchers and clinicians have to account for these variations when reporting Ki67 PIs. To advance the usefulness of Ki67 PIs within the analysis and medical setting, standardization of Ki67 IHC assays is required.
Correct dedication of S-phase fraction in proliferative cells by twin fluorescence and peroxidase immunohistochemistry with 5-bromo-2′-deoxyuridine (BrdU) and Ki67 antibodies.
To make sure the upkeep of tissues in mammals, cell loss should be balanced with cell manufacturing, the proliferative exercise being totally different from tissue to tissue. On this article, the authors suggest a brand new technique for the quantification of the proliferative exercise, outlined because the S-phase fraction of actively biking cells, by twin labeling with fluorescence and peroxidase immunohistochemistry utilizing BrdU (marker of S-phase) and Ki67 antibodies (marker of G(1)-, S-, G(2)-, and M-phases) after a one-step antigen retrieval.
Within the generative cell zones of fundic and pyloric glandular stomachs, the place the vast majority of cells had been biking, the authors measured a proliferative exercise of 31%. Within the epithelium of the forestomach and the pores and skin, the place biking cells are intermingled with G(0) and differentiated cells, proliferative actions had been 21% and 13%, respectively.
Within the adrenal cortex, during which biking cells had been sparsely distributed, the proliferative exercise reached 32%. Through the regenerative course of within the pores and skin after a lesion, the proliferative exercise elevated in proximity to the wound. The current one-step dual-labeling technique has revealed that the proliferative exercise is totally different between tissues and relies on the physiological or pathological state.
Comparability of various Ki67 antibodies in human glioblastomas.
Goals-To check immunostaining between the unique Ki67 monoclonal antibody and a brand new polyclonal Ki67 antibody on frozen and paraffin wax sections of human glioblastomas.Strategies-Frozen sections and formalin fastened, paraffin wax embedded sections of the identical tumour specimens had been included within the examine (10 instances).
Half of the paraffin wax sections had been pretreated in a microwave oven. Customary immunohistochemical methods had been used (avidinbiotin peroxidase complicated). 5 excessive energy fields had been examined utilizing an eye-piece graticule, and 500 to 2000 tumour cells had been counted.
The labelling index was outlined as the proportion of constructive tumour cells.Outcomes-The Ki67 monoclonal antibody displayed constructive immunostaining in all frozen sections (median labelling index 5.9, vary 2.6-11.4) whereas solely 4 paraffin wax sections stained positively and solely after pretreatment in a microwave oven.
The polyclonal Ki67 antibody elicited constructive staining in each frozen sections (median labelling index 13.7, vary 6.7-21.5) and in paraffin wax sections (median labelling index 12.0, vary 2.2-22.7) however solely after pretreatment in a microwave oven.Conclusion-The Ki67 monoclonal antibody just isn’t beneficial to be used on paraffin wax sections of glioma tissues whereas the brand new polyclonal Ki67 antibody gives passable immunostaining on each frozen and paraffin wax sections.
Cell membrane reactivity of MIB-1 antibody to Ki67 in human tumors: truth or artifact?
Ki67 immunohistochemistry is a broadly used marker of the tumor proliferative fraction. Aside from the nuclear staining of dividing cells, MIB-1 monoclonal antibody was additionally discovered to stain the cell membrane of some tumor sorts. Certainly, such membrane reactivity was proposed as a diagnostic characteristic of hyalinizing trabecular tumor (HTT) of the thyroid.
To confirm the diagnostic position of Ki67 membrane sample, 6 HTTs, Eight pulmonary sclerosing hemangiomas (SH), and 6 different human tumors with MIB-1 cell membrane immunoreactivity had been stained by immunoperoxidase with 5 totally different anti-Ki67 antibodies in numerous experimental situations.
We present right here that the cell membrane reactivity reported in HTT is produced solely by MIB-1 and never by different antibodies to Ki67 (together with commercially accessible mouse and rabbit monoclonal antibodies). As well as, this peculiar sample is obtained provided that the response is carried out at room temperature, as a result of automated immunostainers which function at 37 levels C don’t produce any MIB-1 membrane localization. The identical findings had been obtained within the different 6 tumors.
Ki67 Antibody |
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| 48871-50ul | SAB | 50ul | EUR 286.8 |
KI67 antibody |
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| 20R-KR001 | Fitzgerald | 500 ug | EUR 663.6 |
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Description: Rabbit polyclonal KI67 antibody |
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Ki67 Antibody |
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| 33423-100ul | SAB | 100ul | EUR 302.4 |
Ki67 Antibody |
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| 33423-50ul | SAB | 50ul | EUR 224.4 |
KI67 antibody |
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| 10R-K102a | Fitzgerald | 500 ul | EUR 1004.4 |
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Description: Rabbit monoclonal KI67 antibody |
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KI67 antibody |
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| 10R-KR001 | Fitzgerald | 500 ul | EUR 994.8 |
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Description: Rabbit monoclonal KI67 antibody |
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KI67 antibody |
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| 10R-1805 | Fitzgerald | 100 ul | EUR 522 |
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Description: Mouse monoclonal KI67 antibody |
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KI67 antibody |
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| 10R-2147 | Fitzgerald | 100 ul | EUR 483.6 |
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Description: Mouse monoclonal KI67 antibody |
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KI67 antibody |
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| 10R-2244 | Fitzgerald | 100 ul | EUR 457.2 |
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Description: Mouse monoclonal KI67 antibody |
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Ki67 Antibody |
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| 29020-100ul | SAB | 100ul | EUR 302.4 |
Ki67 Antibody |
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| ABF0198 | Lifescience Market | 100 ug | EUR 525.6 |
Ki67 Antibody |
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| ABF5208 | Lifescience Market | 100 ug | EUR 525.6 |
Ki67 Antibody |
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| ABF5498 | Lifescience Market | 100 ug | EUR 525.6 |
KI67 Antibody |
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| BF0132 | Affbiotech | 200ul | EUR 540 |
Ki67 Antibody |
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| DF6818 | Affbiotech | 200ul | EUR 420 |
KI67 Antibody |
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| DF16054 | Affbiotech | 100ul | EUR 420 |
Ki67 Antibody |
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| DF16055 | Affbiotech | 100ul | EUR 420 |
Ki67 Antibody |
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| AF5208 | Affbiotech | 200ul | EUR 420 |
Ki67 Antibody |
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| AF5498 | Affbiotech | 200ul | EUR 420 |
Ki67 Antibody |
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| AF6665 | Affbiotech | 100ul | EUR 420 |
Ki67 Antibody |
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| AF0198 | Affbiotech | 200ul | EUR 420 |
KI67 antibody |
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| PAab09788 | Lifescience Market | 100 ug | EUR 463.2 |
Ki67 Antibody |
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| R20339-0.1ML | NSJ Bioreagents | 100ul | EUR 409 |
Ki67 Antibody |
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| R31458 | NSJ Bioreagents | 100 ug | EUR 419 |
Ki67 antibody |
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| F54499-0.05ML | NSJ Bioreagents | 0.05 ml | EUR 165 |
Ki67 antibody |
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| F54499-0.2ML | NSJ Bioreagents | 0.2 ml | EUR 379 |
Ki67 Antibody |
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| V3864-100UG | NSJ Bioreagents | 100 ug | EUR 499 |
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Description: 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 renal and ureter 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 grade II astrocytoma, oligodendroglioma, colon carcinoma, and breast carcinoma. In general, Ki-67 is a good marker of proliferating cell populations. |
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Ki67 Antibody |
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| V3864-20UG | NSJ Bioreagents | 20 ug | EUR 219 |
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Description: 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 renal and ureter 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 grade II astrocytoma, oligodendroglioma, colon carcinoma, and breast carcinoma. In general, Ki-67 is a good marker of proliferating cell populations. |
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Conversely, sclerosing hemangioma of the lung didn’t produce any MIB-1 cell membrane reactivity in our palms. A cross-reactivity of the MIB-1 monoclonal antibody with an epitope expressed on the cell membrane degree (reasonably than an artifact) appears the probably clarification for this discovering, as a result of the immunoreactivity is mostly intense and uniform within the membrane constructive tumors. We conclude that when Ki67 immunohistochemistry is used for diagnostic functions in a suspected HTT, solely MIB-1 clone at room temperature must be employed.
