monoclonal rat anti-brdu primary antibody  (Bio-Rad)

Journal: PLoS ONE

Article Title: A New Method for In Vitro Detection of Bromodeoxyuridine in Serum: A Proof of Concept in a Songbird Species, the Canary

doi: 10.1371/journal.pone.0063692

Figure Lengend Snippet: (A–F) Cultured cells stained for BrdU, after incubation with BrdU at different concentrations (as indicated at the bottom of each image). Images were consistently modified to enhance contrast and remove colour, and are all shown to the same scale. (A) At very low concentrations, very few cells were labelled even weakly. (B–F) As BrdU concentration increased, the proportion of visible cells that were weakly-labelled (w), moderately-labelled (m), or strongly-labelled (s) increased. Every culture plate also contained some unlabelled cells (arrowhead). (G) Total numbers of labelled cells (weakly+moderately+strongly) reached a plateau with BrdU concentrations above 50 µg/mL; note the break in the x-axis scale . At lower concentrations, however, (inset) there was a strong linear relationship between BrdU concentration and the total number of labelled cells (Y = 1.651 X, line forced through [0,0]; confidence interval for slope = 1.403 to 1.898, r = 0.9606, p<0.0001). Dashed lines show 95% confidence interval around the (solid) line of best fit.

Article Snippet: Cells were then incubated with 0.6% hydrogen peroxide in PBS, rinsed, incubated 10 min in 2N HCl followed by 5 min in 0.1 M sodium borate buffer (pH 8.5), rinsed again, and blocked 8 min in 10% normal horse serum in PBS with 0.02% Triton-X100 (PBS-T) followed by incubation for 2 h at 4°C with monoclonal rat anti-BrdU primary antibody (AbD Serotec, 1∶1000) in PBS-T. Plates were rinsed three times in PBS, followed by incubation for 30 min at room temperature in donkey anti-rat biotinylated secondary antibodies (Jackson Immunoresearch, 1∶2000) in PBS-T. After another three rinses in PBS, plates were incubated 30 min at room temperature with an avidin-biotin complex (Vectastain Elite kit, Vector Laboratories).

Techniques: Cell Culture, Staining, Incubation, Modification, Concentration Assay

Journal: PLoS ONE

Article Title: A New Method for In Vitro Detection of Bromodeoxyuridine in Serum: A Proof of Concept in a Songbird Species, the Canary

doi: 10.1371/journal.pone.0063692

Figure Lengend Snippet: (A). Percentage (means ± SD) of the total numbers of cells visible in culture that were immunoreactive for BrdU after incubation with serum collected from adult canaries at various times after BrdU injection. All individual male (black spots) and female (open circles) data points are represented in the figure. (B) Average concentrations (means ± SD) of BrdU in these canary samples based on the calibration curve illustrated in the inset of . (C–D) Concentrations of BrdU in blood samples collected in two female Japanese quail (C) or one male mouse (D) at various times after a single BrdU injection.

Article Snippet: Cells were then incubated with 0.6% hydrogen peroxide in PBS, rinsed, incubated 10 min in 2N HCl followed by 5 min in 0.1 M sodium borate buffer (pH 8.5), rinsed again, and blocked 8 min in 10% normal horse serum in PBS with 0.02% Triton-X100 (PBS-T) followed by incubation for 2 h at 4°C with monoclonal rat anti-BrdU primary antibody (AbD Serotec, 1∶1000) in PBS-T. Plates were rinsed three times in PBS, followed by incubation for 30 min at room temperature in donkey anti-rat biotinylated secondary antibodies (Jackson Immunoresearch, 1∶2000) in PBS-T. After another three rinses in PBS, plates were incubated 30 min at room temperature with an avidin-biotin complex (Vectastain Elite kit, Vector Laboratories).

Techniques: Incubation, Injection

Journal: Anesthesiology

Article Title: Dantrolene ameliorates impaired neurogenesis and synaptogenesis in Induced Pluripotent Stem Cell lines derived from patients with Alzheimer’s disease

doi: 10.1097/ALN.0000000000003224

Figure Lengend Snippet: (A) Treatment of induced pluripotent stem cells with dantrolene (DAN) (30 μM) for 24 h did not affect induced pluripotent stem cells from healthy human subjects (CON) but resulted in a significantly greater cell viability of induced pluripotent stem cells from sporadic Alzheimer’s disease (SAD) (p=0.006) and familial Alzheimer disease (FAD) (p<0.0001) patients. For cell viability, interaction, treatment and cell type were all significant sources of variation (F(2,40)=92.56, p<0.0001; F(1,40)=110.40, p<0.0001; F(2,40)=92.81, p<0.0001, respectively). (B) Dantrolene proliferation, measured by the percent BrdU positive cells, was significantly impaired in familial Alzheimer’s disease cells compared to control healthy subjects cells (p=0.022). Compared with vehicle control, dimethyl sulfoxide (DMSO), dantrolene resulted in a greater proliferation in familial Alzheimer’s disease cells (p=0.008, familial Alzheimer’s disease dantrolene to dimethyl sulfoxide). For proliferation, dantrolene treatment and cell type were significant sources of variation (F(2,30)=5.44, p=0.009; F(1,30)=9.81, p<0.039, respectively). All data are expressed as the mean±SD from 5–8 independent experiments (A, familial Alzheimer’s disease, n=7; control, n=8; sporadic Alzheimer’s disease, n=8), (B, control treated with DMSO n=7, with dantrolene n=5; sporadic Alzheimer’s disease both DMSO and dantrolene group n=5; familial Alzheimer’s disease DMSO n=8, dantrolene n=6). ** P<0.01, ***P<0.001. Statistical significance was determined using two-way ANOVA with Sidak’s multiple comparison tests.

Article Snippet: After being incubated with blocking solution (5% normal goat serum in phosphate buffered saline containing 0.1% Triton X-100), cells were incubated with rat monoclonal anti-BrdU primary antibody (1:100; Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4°C.

Techniques: Control, Comparison

Journal: Frontiers in Neuroanatomy

Article Title: Downregulation of TrkC Receptors Increases Dendritic Arborization of Purkinje Cells in the Developing Cerebellum of the Opossum, Monodelphis domestica

doi: 10.3389/fnana.2020.00056

Figure Lengend Snippet: Schematic representation of the four main groups of opossums injected with BrdU. (A,C) Opossums of different ages were administered 20 mg/kg body weight of BrdU and perfused at P90 (A) or P35 (C) . (B) Opossums at P35, P50, and P60 were treated with a single dose of BrdU (50 mg/kg) and perfused at P90. (D) Opossums at P35, P60, P90, and P155 were injected with BrdU twice (50 mg/kg). Two hours later, after the second injection of BrdU, opossums were perfused. Each age group consisted of 3–6 opossums.

Article Snippet: After rinsing in TBS, TBS-A, and TBS-B for 10 min each, the sections were incubated in 10% normal goat serum (NGS) solution in TBS-B for 60 min, and then incubated overnight at 4°C with rat anti-BrdU monoclonal primary antibody (1:500, Santa Cruz) in TBS-B.

Techniques: Injection

Journal: Frontiers in Neuroanatomy

Article Title: Downregulation of TrkC Receptors Increases Dendritic Arborization of Purkinje Cells in the Developing Cerebellum of the Opossum, Monodelphis domestica

doi: 10.3389/fnana.2020.00056

Figure Lengend Snippet: Postnatal development of the cerebellum in the Monodelphis opossum. (A–D) Nissl-stained sections in the cerebellum of 2-day-old (A,B) and 5-day-old (C,D) opossums. (E) A coronal section at the level of the cerebellum in an opossum that was injected with BrdU at P1. High-magnification images showing cells of the DCN (F) and Purkinje cells (G) generated on P1 labeled with BrdU. (H–K) Cells labeled with CB (H) , DAPI (I) , BrdU (J) , and merged image of the three labels (K) . (L,M) CB-labeled cells in the cerebellum of a 3-day-old opossum. (N,O) Tbr1-labeled cells in the cerebellum of a 3-day-old opossum. (P–R) CB labeling in the cerebellum of a 35-day-old opossum. IV, fourth ventricle; AQ, aqueduct of Sylvius; Cb, cerebellum; CB, calbindin; CP, choroid plexus; DCN, deep cerebellar nuclei; EGL, external germinal layer; N, neurons; NE, neuroepithelium.

Article Snippet: After rinsing in TBS, TBS-A, and TBS-B for 10 min each, the sections were incubated in 10% normal goat serum (NGS) solution in TBS-B for 60 min, and then incubated overnight at 4°C with rat anti-BrdU monoclonal primary antibody (1:500, Santa Cruz) in TBS-B.

Techniques: Staining, Injection, Generated, Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Downregulation of TrkC Receptors Increases Dendritic Arborization of Purkinje Cells in the Developing Cerebellum of the Opossum, Monodelphis domestica

doi: 10.3389/fnana.2020.00056

Figure Lengend Snippet: Postnatal development of the cerebellum in the opossum. (A–D) Nissl-stained coronal sections on P11 (A,B) and P19 (C,D) opossum brains. (E) Olig2-labeled cells in the cerebellum of an 11-day-old opossum. (F,G) Tbr1-labeled cells in the cerebellum of an 11-day-old opossum. (H,I) BrdU-labeling pattern in the cerebellum of a 3-month-old opossum that was injected with BrdU at P19. (J–L) Double labeling for BrdU (green), Olig2 (red), and merged image of BrdU/Olig2 in the cerebellum of an opossum that was injected with BrdU at P25. (M–O) High-magnification images of double labeling for BrdU (green) and GFAP (red) in the cerebellum of an opossum that was treated with BrdU on P25. arb, arbor vitae; CP, choroid plexus; DCN, deep cerebellar nuclei; EGL, external germinal layer; P, Purkinje cells.

Article Snippet: After rinsing in TBS, TBS-A, and TBS-B for 10 min each, the sections were incubated in 10% normal goat serum (NGS) solution in TBS-B for 60 min, and then incubated overnight at 4°C with rat anti-BrdU monoclonal primary antibody (1:500, Santa Cruz) in TBS-B.

Techniques: Staining, Labeling, Injection

Journal: Frontiers in Neuroanatomy

Article Title: Downregulation of TrkC Receptors Increases Dendritic Arborization of Purkinje Cells in the Developing Cerebellum of the Opossum, Monodelphis domestica

doi: 10.3389/fnana.2020.00056

Figure Lengend Snippet: Proliferative activity in the developing cerebellum of the opossum. (A–D) Immunocytochemical staining for BrdU (green) and GFAP labeling (red) in the cerebellum of P35 (A) , P60 (B) , P90 (C) , and P155 (D) opossums. (E–G) High-magnification confocal images for NeuN (red), BrdU (green), and merged NeuN/BrdU labeling in the cerebellum of a 3-month-old opossum that was injected with BrdU on P60. (H–J) High-magnification confocal images for GFAP (red), BrdU (green), and merged GFAP/BrdU labeling in the cerebellum of a 3-month-old opossum that was injected with BrdU on P60.

Article Snippet: After rinsing in TBS, TBS-A, and TBS-B for 10 min each, the sections were incubated in 10% normal goat serum (NGS) solution in TBS-B for 60 min, and then incubated overnight at 4°C with rat anti-BrdU monoclonal primary antibody (1:500, Santa Cruz) in TBS-B.

Techniques: Activity Assay, Staining, Labeling, Injection