BrdU Monoclonal Antibody (MoBU-1), Pacific Blue
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FIG 3 Cytarabine induces cell cycle arrest and apoptosis of primary effusion lymphoma cells. (A) Effect of cytarabine on cell cycle progression of BJAB, BCBL1, and BC3 cells. Cells treated with 5 uM cytarabine for 48 h were analyzed by flow cytometry after BrdU and propidium iodide staining. (B) Induction of apoptosis in BJAB, BCBL1, and BC3 cells by cytarabine. Cells treated with 5 uM cytarabine for 48 h were analyzed by flow cytometry after annexin 5 and DAPI staining. Early apoptotic cells were those that were positive for only annexin 5, late apoptotic cells were those that were positive for only DAPI, and necrotic cells were those that were positive for both annexin 5 and DAPI. (C) Analysis of apoptosis markers cleaved PARP1 (c-PARP1) and cleaved caspase 3 (c-caspase 3) in BJAB, BCBL1, BC3, BC1, JSC1, and BCP1 cells treated with 5 uM cytarabine for 0, 24, and 48 h by Western blotting.
Figure 3 SIRT1 knockdown or knockout induces cell cycle arrest but has minimal effect on apoptosis in KSHV-transformed cells A. MM and KMM cells with SIRT1 knockdown as described in Figure 2 were pulsed with 10 muM BrdU for 2 h, stained with a pacific blue-conjugated anti-BrdU antibody and propidium iodide, and analyzed by FACS for cell cycle progression. B. MM and KMM cells with SIRT1 knockdown were stained with an anti-Annexin V antibody and analyzed by FACS for Annexin V-positive cells. C. Cell cycle analysis of wild-type and SIRT1 knockout cultures of MM and KMM cells as described in (A). D. Analysis of Annexin V-positive cells in wild-type and SIRT1 knockout cultures of MM and KMM cells as described in (B). Statistical analysis * p <0.05; ** p <0.01; *** p <0.001, NS, not significant.
Published figure using BrdU monoclonal antibody (Product # B35129)
Fig 6 The NF-kappaB pathway mediates KSHV suppression of GLUT1 and GLUT3, and aerobic glycolysis. ( A-B ). Inhibition of the NF-kappaB pathway increases the expression of GLUT1 and GLUT3. MM and KMM cells were treated with NF-kappaB inhibitors JSH23 (30 muM) or BAY 11-7082 (2 muM), and examined for the expression of GLUT1 and GLUT3 expression by RT-qPCR at 8 h post-treatment ( A ) and flow cytometry at 24 h post-treatment ( B ). Y-axis in ( B ) is shown as normalized cell numbers. beta-actin was used as internal controls for qPCR. ( C-D ). Inhibition of the NF-kappaB pathway increases glucose consumption ( C ) and lactate production ( D ). MM and KMM cells were treated with NF-kappaB inhibitor JSH23 (30 muM) and glucose consumption and lactate production were determined as described in Fig 1C and 1E . ( E-F ). Inhibition of the NF-kappaB pathway reduces cell proliferation and inhibits cellular transformation. Cell proliferation ( E ) and colony formation in softagar ( F ) were examined in the presence of JSH23 (30 muM) as described in Fig 1A and 1B . ( G-H ). Inhibition of the NF-kappaB pathway increases apoptosis and reduces BrdU incorporation. Apoptosis ( G ) and BrdU incorporation ( H ) of MM and KMM cells were examined following treatment of JSH23 (30 muM) for 48 h as described in Fig 2D and 2E . All data are presented as mean +- s.e.m. from three (n = 3, A , and C-H ) independent experiments, each with three repeats. Representative images from three independent experimen
Jurkat T-cell leukemia cells were treated with 10µM BrdU for 1.5 hours. The cells were then fixed in ETOH and stored overnight at ≤-20°C. An acid denaturation method was used to prepare the cells before labeling with BrdU mouse monoclonal antibody (Clone MoBU-1) Pacific™ Blue *for flow cytometry* *100 tests* to detect the incorporated BrdU, each using 5 µL antibody conjugate labeling 1 x 10^6 cells. Proliferating cells are clearly distinguished from non-proliferating cells.
Flow Cytometry (Flow)
Host / Isotype
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PBS, pH 7.2
5mM sodium azide
Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine. BrdU is commonly used in the detection of proliferating cells in living tissues, and can be incorporated into the newly synthesized DNA of replicating cells (during the S phase of the cell cycle), substituting for thymidine during DNA replication. Antibodies specific for BrdU can then be used to detect the incorporated chemical, thus indicating cells that were actively replicating their DNA. Binding of the antibody requires denaturation of the DNA, usually by exposing the cells to acid or heat.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.