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Anti-Ki-67 (MRQ-64) is a Reliable Rabbit Monoclonal IHC ...

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Academia Background The Ki-67 antigen is a nuclear, non-histone protein that is present in proliferating cells. Anti-Ki-67 labeling index is the WHO recommended method for grading neoplasms including: glioma, breast carcinoma, lymphoma, sarcoma, and others. 1-5 In this experiment, MRQ-64, a novel rabbit monoclonal antibody, was compared to other commercially available clones. Using an autostainer, immunohistochemistry (IHC) was performed on a variety of tissues including carcinomas, melanomas, sarcomas, and gliomas. Design 44 formalin-fixed paraffin-embedded tissues of various types were selected. All tissues were stained with anti- Ki-67 rabbit monoclonals MRQ-64 and SP6, and mouse monoclonal MIB-1 using an autostainer. Each sample was microscopically evaluated by a pathologist and given a staining score of both signal and background using the following 0-4 scale: 0 = negative, 0.5-2.5 = low intensity and 3-4 = moderate to high intensity. Results All three clones displayed similar sensitivity and intensity, however, it was noted that the background staining intensity with MRQ-64 was almost non-existent providing an excellent overall sensitivity. During this study, pathologist review indicated that the MIB-1 and SP6 clones produced non-specific staining in septic cells, whereas MRQ-64 did not. In total, 12/44 (27.3%) of tissues stained with SP6 and 6/44 (13.6%) of tissues stained with MIB-1 produced noticeable background staining. Furthermore, the MRQ-64 achieved similar scores to the other two clones with more intensity noted on the mitotic figures. Anti-Ki-67 (MRQ-64) is a Reliable Rabbit Monoclonal IHC Marker for Grading Neoplasms Brooke Bivens, B.S.; Qin Su, Ph.D., M.D. Cell Marque™ Tissue Diagnostics, Rocklin, California References: Mckeever P, et al. MIB-1 proliferation index predicts survival among patients with grade II astrocytoma. J Neuropathol Exp Neurol. 1998:57:931-6. Coons SW, et al. The prognostic significance of Ki-67 labeling indices for oligodendrogliomas. Neurosurgery. 1997;41:878-84. Allegra CJ, et al. Prognostic value of thymidylate synthase, Ki-67, and p53 in patients with Dukes’ B and C colon cancer: a National Cancer Institute-national surgical adjuvant breast and bowel project collaborative study. J Clin Oncol. 2003;21:241-50. Pathmanathan N, et al. Ki-67 and proliferation in breast cancer. J Clin Pathol. 2013; 66: 512-6. Swerdlow S, et al. WHO clarifications of tumours of hematopoietic and lymphoid tissues. 4th edition, 2008. P 220. Conclusion Overall, this study demonstrates that anti-Ki-67 (MRQ-64) has equivocal sensitivity and greater specificity when compared with clones SP6 and MIB-1. Background Scores 100 80 60 40 20 0 MIB-1 86.4 13.6 SP6 72.7 27.3 100 MRQ-64 % slides with no background % slides with background The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.
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Page 1: Anti-Ki-67 (MRQ-64) is a Reliable Rabbit Monoclonal IHC ...

Academia

BackgroundThe Ki-67 antigen is a nuclear, non-histone protein that is present in proliferating cells. Anti-Ki-67 labeling index is the WHO recommended method for grading neoplasms including: glioma, breast carcinoma, lymphoma, sarcoma, and others.1-5

In this experiment, MRQ-64, a novel rabbit monoclonal antibody, was compared to other commercially available clones. Using an autostainer, immunohistochemistry (IHC) was performed on a variety of tissues including carcinomas, melanomas, sarcomas, and gliomas.

Design44 formalin-fi xed paraffi n-embedded tissues of various types were selected. All tissues were stained with anti-Ki-67 rabbit monoclonals MRQ-64 and SP6, and mouse monoclonal MIB-1 using an autostainer. Each sample was microscopically evaluated by a pathologist and given a staining score of both signal and background using the following 0-4 scale: 0 = negative, 0.5-2.5 = low intensity and 3-4 = moderate to high intensity.

ResultsAll three clones displayed similar sensitivity and intensity, however, it was noted that the background staining intensity with MRQ-64 was almost non-existent providing an excellent overall sensitivity. During this study, pathologist review indicated that the MIB-1 and SP6 clones produced non-specifi c staining in septic cells, whereas MRQ-64 did not. In total, 12/44 (27.3%) of tissues stained with SP6 and 6/44 (13.6%) of tissues stained with MIB-1 produced noticeable background staining. Furthermore, the MRQ-64 achieved similar scores to the other two clones with more intensity noted on the mitotic fi gures.

Anti-Ki-67 (MRQ-64) is a Reliable Rabbit Monoclonal IHC Marker for Grading NeoplasmsBrooke Bivens, B.S.; Qin Su, Ph.D., M.D.Cell Marque™ Tissue Diagnostics, Rocklin, California

References:

Mckeever P, et al. MIB-1 proliferation index predicts survival among patients with grade II astrocytoma. J Neuropathol Exp Neurol. 1998:57:931-6.

Coons SW, et al. The prognostic signifi cance of Ki-67 labeling indices for oligodendrogliomas. Neurosurgery. 1997;41:878-84.

Allegra CJ, et al. Prognostic value of thymidylate synthase, Ki-67, and p53 in patients with Dukes’ B and C colon cancer: a National Cancer Institute-national surgical adjuvant breast and bowel project collaborative study. J Clin Oncol. 2003;21:241-50.

Pathmanathan N, et al. Ki-67 and proliferation in breast cancer. J Clin Pathol. 2013; 66: 512-6.

Swerdlow S, et al. WHO clarifi cations of tumours of hematopoietic and lymphoid tissues. 4th edition, 2008. P 220.

ConclusionOverall, this study demonstrates that anti-Ki-67 (MRQ-64) has equivocal sensitivity and greater specifi city when compared with clones SP6 and MIB-1.

Background Scores100

80

60

40

20

0MIB-1

86.413.6

SP6

72.7

27.3 100

MRQ-64

% slides with no background

% slides with background

The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.

Page 2: Anti-Ki-67 (MRQ-64) is a Reliable Rabbit Monoclonal IHC ...

Figure 2A: Germinal center of tonsil tissue stained with anti-Ki-67 (SP6)

Figure 1A: Germinal center of tonsil tissue stained with anti-Ki-67 (MIB-1)

Figure 2B: Infl ammatory cells of tonsil tissue stained with anti-Ki-67 (SP6)

Figure 1B: Infl ammatory cells of tonsil tissue stained with anti-Ki-67 (MIB-1)

Figure 3A: Germinal center of tonsil tissue stained with anti-Ki-67 (MRQ-64)

Figure 3B: Infl ammatory cells of tonsil tissue produced no stain with anti-Ki-67 (MRQ-64)

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