New antiserum against Ki-67 antigen suitable for double immunostaining of paraffin wax sections.

Key, Göran; Petersen, J.; Becker, M. H. G.; Duchrow, M.; Schlüter, Carsten; Askaa, Jon; Gerdes, Johannes

doi:10.1136/jcp.46.12.1080

Cited by 81 publications

(44 citation statements)

References 17 publications

(1 reference statement)

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“…One-month-old mice were injected with BrdU (six injections over 48 hours -the pulse period) to label replicating cells, and sacrificed at one of several subsequent time points (the chase period). Beta cells that replicated during the pulse period should stain for BrdU, whereas beta cells that were dividing at the time of sacrifice should stain for the general cell cycle marker Ki67 (Gerdes et al, 1991;Key et al, 1993). The fraction of recently replicated beta cells that re-entered the cell cycle can be determined by staining beta cells for BrdU and Ki67 at the chase time periods and calculating the percentage of BrdU + Ki67 + cells among the BrdU-labeled population (Fig.…”

Section: Resultsmentioning

confidence: 99%

Glucose and aging control the quiescence period that follows pancreatic beta cell replication

et al. 2010

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SUMMARYPancreatic beta cell proliferation has emerged as the principal mechanism for homeostatic maintenance of beta cell mass during adult life. This underscores the importance of understanding the mechanisms of beta cell replication and suggests novel approaches for regenerative therapy to treat diabetes. Here we use an in vivo pulse-chase labeling assay to investigate the replication dynamics of adult mouse beta cells. We find that replicated beta cells are able to re-enter the cell division cycle shortly after mitosis and regain their normal proliferative potential after a short quiescence period of several days. This quiescence period is lengthened with advanced age, but shortened during injury-driven beta cell regeneration and following treatment with a pharmacological activator of glucokinase, providing strong evidence that metabolic demand is a key determinant of cell cycle re-entry. Lastly, we show that cyclin D2, a crucial factor in beta cell replication, is downregulated during cell division, and is slowly upregulated post-mitosis by a glucose-sensitive mechanism. These results demonstrate that beta cells quickly regain their capacity to re-enter the cell cycle post-mitosis and implicate glucose control of cyclin D2 expression in the regulation of this process.

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Section: Resultsmentioning

confidence: 99%

Glucose and aging control the quiescence period that follows pancreatic beta cell replication

et al. 2010

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“…The sheep Tbr2 antibody was characterized by western blot on lysates of BG01V, mesendoderm‐differentiated human embryonic stem cells, where it recognized specific bands at 85–105 kDa. The Ki67 antibody was characterized by western blot of IM‐9 (multiple myeloma cell line) cell lysates, where it detected bands of 345 and 395 kDa, identical to the original Ki‐67 antibody (manufacturer's information; Key et al, 1993). Additionally, the pattern of Ki67 staining we observed was similar to that of previous ferret studies using a different antibody (Reillo and Borrell, 2012).…”

Section: Methodsmentioning

confidence: 99%

S‐phase duration is the main target of cell cycle regulation in neural progenitors of developing ferret neocortex

García

Chang

Arai

et al. 2015

J of Comparative Neurology

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The evolutionary expansion of the neocortex primarily reflects increases in abundance and proliferative capacity of cortical progenitors and in the length of the neurogenic period during development. Cell cycle parameters of neocortical progenitors are an important determinant of cortical development. The ferret (Mustela putorius furo), a gyrencephalic mammal, has gained increasing importance as a model for studying corticogenesis. Here, we have studied the abundance, proliferation, and cell cycle parameters of different neural progenitor types, defined by their differential expression of the transcription factors Pax6 and Tbr2, in the various germinal zones of developing ferret neocortex. We focused our analyses on postnatal day 1, a late stage of cortical neurogenesis when upper‐layer neurons are produced. Based on cumulative 5‐ethynyl‐2′‐deoxyuridine (EdU) labeling as well as Ki67 and proliferating cell nuclear antigen (PCNA) immunofluorescence, we determined the duration of the various cell cycle phases of the different neocortical progenitor subpopulations. Ferret neocortical progenitors were found to exhibit longer cell cycles than those of rodents and little variation in the duration of G1 among distinct progenitor types, also in contrast to rodents. Remarkably, the main difference in cell cycle parameters among the various progenitor types was the duration of S‐phase, which became shorter as progenitors progressively changed transcription factor expression from patterns characteristic of self‐renewal to those of neuron production. Hence, S‐phase duration emerges as major target of cell cycle regulation in cortical progenitors of this gyrencephalic mammal. J. Comp. Neurol. 524:456–470, 2016. © 2015 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc.

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“…It would be prudent when monitoring cell numbers over time, however, to use one sample type alone in order to minimise the variables affecting the results obtained. 6) was used for washes and for diluting the antibodies with the exception of the primary antibodies, to which 10% normal swine serum was added. Sections were placed on coated slides (Fro-tissuer pen, The Binding Site) and after incubation and subsequent rehydration, were pretreated in a microwave oven according to standard protocols.8 Sections were immunostained using the Dako Duet Strept ABC HRP system (K492, Dako), but a biotinylated rabbit anti-goat secondary antibody (E466, Dako), at a 1 in 100 dilution, was applied for the detection of sheep polyclonal Ki67.…”

Section: Discussionmentioning

confidence: 99%

Detection of Ki67 antigen by a new sheep polyclonal antiserum.

Reynolds

Rowlands

Mead

1995

Journal of Clinical Pathology

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Results Samples were obtained from 22 children aged from eight months to 16-4 years, median 5-5 years, and 10 adults aged between 26 and 44 years. Data from adults and children were combined for analysis. The numbers of the different cell types were higher in skin puncture blood (4-8-6-4% higher for subsets other than CD3+ CD16+ CD56+ natural killer cells, which were 10-7% higher), the difference being statistically significant for all cell types except CD3+ CD8+ T lymphocytes (table). Significance was lost for all pairings when total lymphocytes and subpopulations were expressed in proportional terms (data not shown). Paired values were highly correlated (r=0 90 for CD3 + CD16 + CD56 + natural killer cells, and 0-96-0.98 for total white cells, lymphocytes and other subsets, p<00001 for all pairings). The differences between paired CD3 + CD4 + T cell counts is shown in the figure. DiscussionCollection of blood by skin puncture provides a rapid and simple altemative to venepuncture when small volumes of blood are required, and is useful when dealing with small children and subjects with difficult veins. In this study no undue difficulty was found in obtaining the required volume ofskin puncture blood; should problems arise it may be possible to reduce test volumes by 25-50%, at least when the total lymphocyte count is within normal limits. Previous studies have shown clinically acceptable comparability in white cell and differential counts between the two samples types,2 3 although in the earlier study neutrophil values were significantly higher in skin puncture blood. Similar results were obtained in the present study, the usually slightly higher total white cell count of skin puncture samples being reflected proportionally throughout the range of lymphocyte subsets. Although of borderline statistical significance for most subpopulations, the magnitude of the difference was small and unlikely to be of clinical importance.The precision of both replicate haemoglobin and platelet estimations from skin puncture samples has been reported to be poorer than that from venous blood,45 although the latter claim could not be verified using the same sample collection method as that of the present study.2 While we did not count lymphocytes in sequential paired samples there appears to be no reason why their numbers should fluctuate to a greater degree in skin puncture blood, a contention supported by the highly significant correlations found for each lymphocyte subpopulation between the two sample types.Our results indicate that skin puncture blood can be safely used as an alternative to venous blood for the enumeration of CD4 and other lymphocyte subsets. It would be prudent when monitoring cell numbers over time, however, to use one sample type alone in order to minimise the variables affecting the results obtained.1 Calvelli T, Denny TN, Paxton H, Galman R, Kagan J.Guidelines for flow cytometric immunophenotyping: a report from the National Institute of Allergy and Infectious Diseases, Division of AIDS. Cytometry 1993;14:...

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New antiserum against Ki-67 antigen suitable for double immunostaining of paraffin wax sections.

Cited by 81 publications

References 17 publications

Glucose and aging control the quiescence period that follows pancreatic beta cell replication

Glucose and aging control the quiescence period that follows pancreatic beta cell replication

S‐phase duration is the main target of cell cycle regulation in neural progenitors of developing ferret neocortex

Detection of Ki67 antigen by a new sheep polyclonal antiserum.

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