The cells were seeded and serum-starved, as described for the metabolic activity assay. Then, the cells were treated with different concentrations of CSE with or without 10 µM L-NIL, for 6 h. The medium was replaced with a fresh medium containing BrdU labeling solution (Roche, Basel, Switzerland), and the cells were cultured for an additional 16 h. For the experiments with BALF, a serum-free medium containing BrdU labeling solution (Roche, Basel, Switzerland) and either BALF diluted 1:10 or PBS (as a control) was added, and the cells were cultured for 16 h. Next, the cells were fixed and proliferation was determined, in accordance with the manufacturer’s instructions. The absorbance was measured in a microplate reader at 370 nm (reference wavelength: 492 nm). The O.D. value for each experimental sample was standardized to the value measured in the control group.
Brdu labeling solution
Manufactured by Roche
Sourced in Germany
BrdU labeling solution is a reagent used in cell proliferation assays. It contains bromodeoxyuridine (BrdU), a synthetic nucleoside that can be incorporated into the newly synthesized DNA of replicating cells. This enables the detection and quantification of cell division.
Automatically generated - may contain errors
Lab products found in correlation
Ly364947, by Merck Group (1 mentions)
Noggin, by R&D Systems (1 mentions)
Bone morphogenetic protein 4 (bmp4), by R&D Systems (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Roche (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Model 550, by Bio-Rad (1 mentions)
Ix50 microscope, by Olympus (1 mentions)
Elisa reader, by Tecan (1 mentions)
Brdu antibody, by Abcam (1 mentions)
H 2kd sf1 1, by Thermo Fisher Scientific (1 mentions)
H2 db 28 14 8, by BD (1 mentions)
Macs microbead, by Miltenyi Biotec (1 mentions)
7 protocols using brdu labeling solution
1
Cell Proliferation Assay with CSE
2
Directed Differentiation of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were treated with 20 ng/mL BMP4 (R&D System), 250 ng/mL noggin (R&D System), 20 ng/mL TGF-β1 (R&D System), 3 μM LY364947 (Sigma), and 10 μM BrdU labeling solution (Roche), as described.
3
Lymphocyte Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single-cell suspensions of lymph nodes and spleen of mice were prepared under sterile conditions, on day 25 post-immunization. Primed lymphoid populations passed through a 40 μm cell strainer followed by ammonium chloride-based erythrocyte lysis (BD Biosciences, Heidelberg, Germany). Subsequently, single-cell suspensions were prepared in a complete medium consisting of RPMI-1640 medium, 10% fetal bovine serum (FBS), 100 IU penicillin/mL, and 100 μL streptomycin/mL (all reagents given by Sigma, St. Louis, MO, USA). Cells were then located into round-bottom 24-well and 96-well plates respectively 2 × 106 cells and 2 × 105 cells to each well and incubated for 72 h at 37 °C and 5% CO2. To stimulate the cells, MOG35–55 peptide (20 μg/mL) and PHA (20 μg/mL) were added. Throughout the last 24 h, the cells were cultured in the presence of a BrdU-labeling solution (Roche Applied Sciences, Penzberg, Germany). Afterward, proliferation was measured using the Roche Cell Proliferation ELISA BrdU kit, conferring to the manufacturer guidelines. The final steps of this test were picked up in a Stat Fax 2100 Awareness microplate reader (Fisher BioBlock Scientific, Palm City, FL, USA) at 450 nm.
4
BrdU Proliferation Assay for HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bromodeoxyuridine (BrdU; (Roche, Basel, Switzerland) assay was performed as described previously (15 (link)). HUVECs incubated with different types of HDL at 100 µg/ml apoA-I concentration for 12, 24, 36 or 48 h. The cells were labeled with BrdU labeling solution (Roche) and then fixed with paraformaldehyde (Sigma-Aldrich). Following incubation with peroxidase-conjugated anti-BrdU working solution (Roche) for 90 min at 37°C, the cells were washed with washing buffer three times and substrate solution, 3,3′,5,5′-tetramethylbenzidine (Roche) was added. The absorbance of each well was measured at a wavelength of 450 nm with an ELISA plate reader (Model 550; Bio-Rad Laboratories, Inc., Hercules, CA, USA).
5
Pericyte-Mediated Angiogenesis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs were labeled by BrdU labeling solution (Roche) and treated with conditioned medium for 24 h. The level of incorporated BrdU was measured at OD370 with an ELISA reader (TECAN). To measure cell migration, HUVECs (20,000 cells) were seeded into the lower transwell with the plastic insert (ibidi) and cultured overnight. Pericytes were infected with adenoviral vector expressing Sema3C or GFP as a control and grown on transwells for 24 h. A gap within the HUVEC monolayer was generated by removing the inserts, and the growth medium was replaced with Endopan3 basal medium (0.5% FCS). At the same time, pericytes were cocultured with HUVEC. The gap in the HUVEC monolayer was imaged with an Olympus IX 50 microscope at 4× magnification at 0 and 24 h. Bright-field images were taken and analyzed with Cell^P software. Apoptosis was measured with the Caspase-Glo®3/7 Substrate solution (Sigma) after 3 h of incubation of conditioned medium.
6
BrdU Incorporation Assay Optimization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in 6-well plates (3×105 cells/well). After BIX-01294 treatment, BrdU labeling solution (Roche, Palo Alto, CA) was added into each well for 1 h at 37°C. After fixation by 2% paraformaldehyde for 15 min at room temperature and PBS washing for 3 times, cells were incubated with BrdU antibody (Abcam, 1:250) for 1 h at 37°C. Cells were then washed with PBS and incubated with alkaline phosophatase (AP) conjugated secondary antibody (Abcam) at room temperature for 16 h. After PBS washing, the cells were treated with colorsubstrate solution (Pierce, IL, USA) at room temperature for 20 min. The stained cells were photographed and 5 randomly chosen visual fields were counted to determine the percentage of BrdU positive cells.
7
Insulin-Loaded DC Stimulate NOD CD8+ T Cells
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!