Key Specifications Table
|Species Reactivity||Key Applications||Host||Format||Antibody Type|
|B, Ca, H, M, R, Rb, F, Vrt||IF, ICC, IH(P)||M||Ascites||Monoclonal Antibody|
|Description||Anti-PCNA Antibody, clone 131-11912|
|Presentation||Ascites fluid containing no preservatives|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Maintain frozen at -20°C in undiluted aliquots for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles.|
|Material Size||100 µL|
|Reference overview||Pub Med ID|
|Temozolomide PLGA microparticles: a new protocol for treatment of glioma in rats. |
Zhang YH, Zhang H, Liu JM, Yue ZJ
Medical oncology (Northwood, London, England) 28 901-6. Epub 2010 Apr 16. 2011
Implantable and poly (d,l-lactide-co-glycolide) (PLGA) microparticles were developed to deliver temozolomide (TM) continuously in interstitial chemotherapy for glioma. The therapeutic effect of temozolomide/PLGA was evaluated in a rat C6 glioma model. C6 cells were implanted orthotopically into 100 rat brains in 5 groups (n=20 each): sham operation group, control group, local delivery of blank PLGA microspheres group, oral TM group, and local delivery of TM/PLGA group. Rats in oral TM group were orally administered temozolomide, and rats in TM/PLGA group were locally implanted with TM/PLGA microspheres. Ten rats were selected randomly from each group for observing the survival time, and the other 10 rats were killed on POD 14 to measure proliferation activity and apoptosis of the gliomas. Head MRI examination was performed before the rats were killed. The median survival time of sham operation group, control group, blank PLGA microspheres group, oral TM group, and TM/PLGA group was 19.5, 20, 19, 27, and 46.5 days, respectively. MRI demonstrated that the tumor volume was reduced in oral TM group and interstitial TM/PLGA group. PCNA-positive cell staining showed that proliferation activity of tumor cells treated with interstitial TM/PLGA therapy significantly decreased when compared with that of tumor cells treated with oral TM therapy. The apoptosis of C6 cells in interstitial TM/PLGA group significantly increased when compared with that in oral TM group. Interstitial TM/PLGA was effective in treating intracranial C6 rat gliomas and could prove to be a potential chemotherapy agent for human malignant gliomas.
|A COMPARISON OF DIFFERENTIATION PROTOCOLS FOR RGC-5 CELLS. |
Wood JP, Chidlow G, Tran T, Crowston J, Casson RJ
Invest Ophthalmol Vis Sci 2010
Purpose: Although the RGC-5 cell line is widely employed in retinal ganglion cell (RGC) research, recent data have raised questions about the nature of these cells. We therefore performed a systematic analysis of RGC-5 cells in order to determine which RGC or neuronal markers are expressed after treatment with known differentiating agents. This provided further insights into the nature of these cells and assisted in defining their future use. Methods: RGC-5 cells were treated for 5 days with either staurosporine (STSN; 316nM), trichostatin A (TSA; 500nM) or succinyl-concanavalin A (sConA; 50microg/ml), whereafter they were assayed for specific marker antigen/mRNA expression. Treated cells were also assayed for excitotoxic responsiveness. Results: Neither treated nor untreated RGC-5 cells expressed any specific RGC marker mRNAs or proteins (Brn-3, neurofilaments, Thy-1), or any of calbindin, calretinin, synaptophysin, PKCalpha or glial fibrillary acidic protein (GFAP). However, control RGC-5 cells did express the neuronal markers, tau, betaIII-tubulin, MAP-1b, MAP2 and PGP9.5. Although treatment with sConA had no effect on expression of these markers, STSN and TSA (the latter, dose-dependently) increased their expression and induced excitotoxic responsiveness. All cells, whether treated or not, expressed high levels of nestin, but no other progenitor cell markers. All cells also expressed cone-specific but not rod-specific opsin indicative of being of cone photoreceptor lineage. Conclusions: RGC-5 cells expressed neuronal but not RGC-specific markers which were dose-dependently upregulated by TSA. Hence, TSA provided the best tested means to terminally differentiate the cells to a neuronal phenotype from a precursor-like lineage.
|Temozolomide/PLGA microparticles plus vatalanib inhibits tumor growth and angiogenesis in an orthotopic glioma model. |
Yu-Hui Zhang,Zhi-Jian Yue,He Zhang,Gu-Sheng Tang,Yang Wang,Jian-Min Liu
European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft für Pharmazeutische Verfahrenstechnik e.V 76 2010
Temozolomide (TM) has anti-tumor activity in patients with malignant glioma. Implantable poly (D,L-lactide-co-glycolide) (PLGA) microparticles of TM (TM-MS) have been developed, enhancing the cytotoxicity of TM to Glioma C6 cells. Vatalanib, as anti-angiogenic agent, has also shown anti-tumor activity with malignant gliomas. We examined the combined effects of TM-MS and vatalanib in a rat orthotopic glioma model and found TM-MS offered a greater tumor inhibition than TM, and combination treatment with both of them improved the survival time versus single agent therapy. The combination treatment also demonstrated an inhibition to rat glioma tumors, a significant decrease in cell proliferation, an increase in apoptosis, and a lower microvessel density within the glioma tumors. The results suggest that TM-MS can more effectively inhibit tumor than TM, and combination treatment with TM-MS and vatalanib inhibits tumor growth and angiogenesis and may prove to be a promising therapy for malignant gliomas.
|Drosophila CAF-1 regulates HP1-mediated epigenetic silencing and pericentric heterochromatin stability. |
Huang H, Yu Z, Zhang S, Liang X, Chen J, Li C, Ma J, Jiao R
J Cell Sci 123 2853-61. Epub 2010 Jul 27. 2010
Chromatin assembly factor 1 (CAF-1) was initially characterized as a histone deliver in the process of DNA-replication-coupled chromatin assembly in eukaryotic cells. Here, we report that CAF-1 p180, the largest subunit of Drosophila CAF-1, participates in the process of heterochromatin formation and functions to maintain pericentric heterochromatin stability. We provide evidence that Drosophila CAF-1 p180 plays a role in both classes of position effect variegation (PEV) and in the expression of heterochromatic genes. A decrease in the expression of Drosophila CAF-1 p180 leads to a decrease in both H3K9 methylation at pericentric heterochromatin regions and the recruitment of heterochromatin protein 1 (HP1) to the chromocenter of the polytene chromosomes. The artificial targeting of HP1 to a euchromatin location leads to the enrichment of Drosophila CAF-1 p180 at this ectopic heterochromatin, suggesting the mutual recruitment of HP1 and CAF-1 p180. We also show that the spreading of heterochromatin is compromised in flies that have reduced CAF-1 p180. Furthermore, reduced CAF-1 p180 causes a defect in the dynamics of heterochromatic markers in early Drosophila embryos. Together, these findings suggest that Drosophila CAF-1 p180 is an essential factor in the epigenetic control of heterochromatin formation and/or maintenance.
|Decreased neurogenesis in aged rats results from loss of granule cell precursors without lengthening of the cell cycle. |
Ana Olariu,Kathryn M Cleaver,Heather A Cameron
The Journal of comparative neurology 501 2007
It is well established that neurogenesis in the dentate gyrus slows with aging, but it is unclear whether this change is due to slowing of the cell cycle, as occurs during development, or to loss of precursor cells. In the current study, we find that the cell cycle time of granule cell precursors in middle-aged male rats is not significantly different from that in young adults. The size of the precursor pool, however, was 3-4 times smaller in the middle-aged rats, as determined using both cumulative bromodeoxyuridine (BrdU) labeling as well as labeling with the endogenous marker of cell proliferation, proliferating cell nuclear antigen (PCNA). Loss of precursor cells was much greater in the granule cell layer than in the hilus, suggesting that dividing cells in the hilus belong to a distinct population, most likely glial progenitors, that are less affected by aging than neuronal precursors. BrdU-labeled precursor cells and young neurons, labeled with doublecortin, appeared to be lost equally from rostral and caudal, as well as suprapyramidal and infrapyramidal, subregions of the granule cell layer. However, doublecortin staining did show large parts of the caudal granule cell layer with few if any young neurons at both ages. Taken together, these findings indicate that precursor cells are not distributed evenly within the dentate gyrus in adulthood but that precursors are lost from throughout the dentate gyrus in old age with no concomitant change in the cell cycle time.
|Determination of key aspects of precursor cell proliferation, cell cycle length and kinetics in the adult mouse subgranular zone. |
Mandyam, CD; Harburg, GC; Eisch, AJ
Neuroscience 146 108-22 2007
Neurogenesis studies on the adult mouse hippocampal subgranular zone (SGZ) typically report increases or decreases in proliferation. However, key information is lacking about these proliferating SGZ precursors, from the fundamental--what dose of bromodeoxyuridine (BrdU) is appropriate for labeling all S phase cells?--to the detailed--what are the kinetics of BrdU-labeled cells and their progeny? To address these questions, adult C57BL/6J mice were injected with BrdU and BrdU-immunoreactive (IR) cells were quantified. Initial experiments with a range of BrdU doses (25-500 mg/kg) suggested that 150 mg/kg labels all actively dividing precursors in the mouse SGZ. Experiments using a saturating dose of BrdU suggested BrdU bioavailability is less than 15 min, notably shorter than in the developing mouse brain. We next explored precursor division and maturation by tracking the number of BrdU-IR cells and colabeling of BrdU with other cell cycle proteins from 15 min to 30 days after BrdU. We found that BrdU and the Gap2 and mitosis (G2/M) phase protein pHisH3 maximally colocalized 8 h after BrdU, indicating that the mouse SGZ precursor cell cycle length is 14 h. In addition, triple labeling with BrdU and proliferating cell nuclear antigen (PCNA) and Ki-67 showed that BrdU-IR precursors and/or their progeny express these endogenous cell cycle proteins up to 4 days after BrdU injection. However, the proportion of BrdU/Ki-67-IR cells declined at a greater rate than the proportion of BrdU/PCNA-IR cells. This suggests that PCNA protein is detectable long after cell cycle exit, and that reliance on PCNA may overestimate the length of time a cell remains in the cell cycle. These findings will be critical for future studies examining the regulation of SGZ precursor kinetics in adult mice, and hopefully will encourage the field to move beyond counting BrdU-IR cells to a more mechanistic analysis of adult neurogenesis.Full Text Article
|Proliferating cell nuclear antigen immunohistochemistry using monoclonal antibody 19A2 and a new antigen retrieval technique has prognostic impact in archival paraffin-embedded node-negative breast cancer. |
Siitonen, S M, et al.
Am. J. Pathol., 142: 1081-9 (1993) 1993
We evaluated whether proliferating cell nuclear antigen (PCNA) immunohistochemistry with antigen retrieval could be used as a measure of cell proliferation in archival, formalin-fixed, paraffin-embedded tissues and whether the staining results have long-term prognostic significance in axillary node-negative breast cancer. Primary tumor samples obtained from 109 axillary-node-negative breast cancer cases were used for the study. The best staining results were obtained with the 19A2 antibody after microwave heating in a solution of saturated lead thiocyanate. Using this method, there was a significant correlation (linear regression, r = 0.580, P < 0.001) between the proportion of PCNA19A2-positive carcinoma cells (PCNA19A2 score) and DNA flow cytometric S phase fraction. A high PCNA19A2 score was associated with high mitotic count, DNA aneuploidy, and absence of estrogen receptors. Axillary-node-negative patients with a high PCNA19A2 score (cut-point 8%) had significantly worse prognosis than those with a low PCNA19A2 score (P = 0.008). According to a Cox multivariate analysis, PCNA19A2 score had independent prognostic value but only if S phase fraction was excluded from the analysis. In our study, the PCNAPC10 score correlated weakly only with primary tumor size (analysis of variance) and prognosis (5-year univariate survival analysis), but the significance of these findings needs further evaluation. In conclusion, PCNA immunohistochemistry with the 19A2 antibody after an appropriate antigen retrieval treatment may offer a useful alternative to DNA flow cytometry for the analysis of cell proliferation activity from formalin-fixed, paraffin-embedded breast carcinomas.