Brdu solution

Manufactured by Beyotime

Sourced in China

BrdU solution is a laboratory reagent used in biological research. It is a synthetic nucleoside that is an analog of the DNA base thymine. BrdU can be incorporated into the newly synthesized DNA of dividing cells, allowing for the identification and tracking of these cells.

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Lab products found in correlation

Anti brdu antibody, by Beyotime (3 mentions) Ab150113, by Abcam (1 mentions) Goatanti mouse igg h l antibody, by Abcam (1 mentions) Ab8152, by Abcam (1 mentions) Cx41 32rfl, by Olympus (1 mentions) Brdu cell proliferation assay kit, by Cell Signaling Technology (1 mentions) Hoechst staining solution, by Beyotime (1 mentions) Hoechest staining solution, by Beyotime (1 mentions) Alexa fluor 488, by Abcam (1 mentions) Anti brdu primary antibody, by Merck Group (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Dapi staining solution, by Beyotime (1 mentions) Ab220074, by Abcam (1 mentions) Dapi buffer, by Beyotime (1 mentions) Fluorescent microscope, by Olympus (1 mentions)

10 protocols using brdu solution

BrdU Incorporation Assay for Proliferation

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U87/TMZ and U251/TMZ cells were transferred to 24-
well plates. After the cells were cultured in serum-free
medium for 24 hours, the cells were incubated with 10
μmol/L of BrdU solution (Beyotime, Shanghai, China) in
a complete medium for 4 hours at 37°C. After the medium
was removed, the cells were rinsed 3 times with phosphate
buffer saline (PBS), fixed with 70% ethanol for 10 minutes
at 4°C, and then washed 3 times with PBS. Then, 2 mol/L
of HCl was added to denature the cellular DNA at 37°C
for 60 minutes before the cells were washed 3 times with
PBS. After that, the cells were blocked with 3% bovine
serum albumin (KPL, Gaithersburg, MD, USA) at room
temperature for 1 hour, followed by the rinse with PBS 3
times, 5 minutes for each time. Subsequently, the cells were
incubated overnight at 4°C with anti-BrdU monoclonal
antibody (ab8152, 1:300, Abcam, Shanghai, China) and
then with goat anti-mouse fluorescent secondary Goat
Anti-Mouse IgG H&L antibody (ab150113, 1:1000,
Abcam, Shanghai, China) at room temperature for 2 hours.
After counterstaining the cell nucleus with DAPI, the
cells were observed under a fluorescence microscope. In
10 randomly selected non-overlapping fields, the number
of BrdU-positive cells was counted, and the average was
calculated.

Houjun Z, & Peng B. (2023). Down-Regulation of CEND1 Expression Contributes to The Progression and Temozolomide Resistance of Glioma. Cell Journal (Yakhteh), 25(4), 264-272.

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Quantifying Proliferative Capacity of GC Cells

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Following transfection, GC cells in each group were moved onto coverslips (Beyotime, Shanghai, China) for 12 hours of culture. Later, the cells were incubated with BrdU solution (Beyotime, Shanghai, China) for 6 hours. The culture medium was discarded. GC cells were immobilized with 4% paraformaldehyde for 30 minutes, incubated with anti-BrdU antibody (Beyotime, Shanghai, China) for an hour at room temperature (RT), and flushed with phosphate buffer saline (PBS). The number of positive BrdU cells was calculated [25 (link)].

Tan Y., Li Y., Zhu H., Wu X., Mei K., Li P, & Yang Q. (2022). miR-187/PDLIM1 Gets Involved in Gastric Cancer Progression and Cisplatin Sensitivity of Cisplatin by Mediating the Hippo-YAP Signaling Pathway. Journal of Oncology, 2022, 5456016.

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BrdU Cell Proliferation Assay Protocol

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BrdU cell proliferation assay kit (Cell Signaling Technology Inc.) was used for BrdU cell proliferation assay. Transfected C33A and HeLa cells were seeded (5×10³ cells/ml) into 96-well plates and cultured overnight. Subsequently, 20 µl BrdU solution (Beyotime Institute of Biotechnology) was added to each well and incubated for 12 h. Subsequently, the culture medium was discarded and cells were washed with PBS. Cells were fixed with 4% paraformaldehyde for 30 min at room temperature and washed again with PBS. Cells were incubated with anti-BrdU (cat. no. ab6326; 1:2,000; Abcam) for 1 h at room temperature. Then, cell nuclei were counterstained using Hoechst staining solution (Beyotime Institute of Biotechnology) at room temperature for 30 min. BrdU⁺ cells were observed and counted using a fluorescence microscope (CX41-32RFL; Olympus Corporation).

Yang X., Du H., Bian W., Li Q, & Sun H. (2021). FOXD3-AS1/miR-128-3p/LIMK1 axis regulates cervical cancer progression. Oncology Reports, 45(5), 62.

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BrdU Cell Proliferation Assay

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The transfected GC cells in each group were transferred onto coverslips (Beyotime, Shanghai, China) and then cultured for 12 h. Subsequently, the cells were incubated BrdU solution (Beyotime, Shanghai, China) for 6 h. The GC cells, after the medium was discarded, were fixed with 4% paraformaldehyde for 30 min, and incubated with anti-BrdU antibody (Beyotime, Shanghai, China) at ambient temperature for 1 h, and then rinsed with phosphate buffer saline (PBS). Subsequently, the nuclei were stained employing Hoechest staining solution (Beyotime, Shanghai, China). After the cells were washed by PBS again, the numbers of BrdU and Hoechst positive cells were counted.

Tao F., Qi L, & Liu G. (2022). Long intergenic non-protein coding RNA 662 accelerates the progression of gastric cancer through up-regulating centrosomal protein 55 by sponging microRNA-195-5p. Bioengineered, 13(2), 3007-3018.

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Cell Proliferation Assay via BrdU

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MDA-MB-231 and MCF-7 cells were transferred into 24-well plates (with a cover glass in each well, 2.5 × 10⁵ cells/well), respectively. After 24 h, the cells were incubated with BrdU solution (Beyotime Biotechnology, Shanghai, China) for another 4 h and then fixed for 30 min with 4% paraformaldehyde. After the supernatant was discarded, the cells were incubated with anti-BrdU antibody (1: 500, Beyotime Biotechnology, Shanghai, China) for 30 min at room temperature and then with 100 μL of 1× Hoechst 33,342 reaction solution at room temperature in the dark for 20 min, followed by being rinsed with PBS. In 10 random high-power fields under the microscope, the number of BrdU-positive cells and the total number of cells were calculated, and the average was taken.

Wu X., Kong C, & Wu Y. (2021). Long intergenic non-protein coding RNA 1094 (LINC01094) promotes the progression of breast cancer (BC) by regulating the microRNA-340-5p (miR-340-5p)/E2F transcription factor 3 (E2F3) axis. Bioengineered, 12(1), 9046-9057.

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Cell Proliferation Measurement Protocol

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Briefly, LN-229 and T98-G cells were plated in 24-well plates at 2.5 × 10⁴ cells/well, and cultured for 24 h, followed by the addition of BrdU solution (Beyotime Biotechnology, Shanghai, China) to incubate for another 4 h. Then, the cells were fixed, and incubated with anti-BrdU antibody for 2 h, and then incubated with secondary antibodies for 1 h. The nucleus was marked by 10 μmol/L DAPI for 5 min. Ultimately, an upturned fluorescence microscope was utilized for observing the cells, and the percentage of BrdU-positive cells was calculated.

Ma Z., Chen Z., Zhou Y., Li Y., Li S., Wang H, & Feng J. (2022). Hsa_circ_0000418 promotes the progression of glioma by regulating microRNA-409-3p / pyruvate dehydrogenase kinase 1 axis. Bioengineered, 13(3), 7541-7552.

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Cell Proliferation Assay Using BrdU

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HEp-2 and TU212 cells were transferred at 2.5 × 10⁵ cells/well to 24-well plates (with a cover slide in it). BrdU solution (Beyotime Biotechnology, Shanghai, China) was added after 24 h of culture, and the culture was continued for 4 h. Then, the cells were washed with PBS, and fixed for 30 min with 4% cold paraformaldehyde; the supernatant was discarded and 1× Apollo (100 μL) was added to each well for cell staining for 30 min; the supernatant was discarded and each well was added with 1× Hoechst 33,342 reaction solution (100 μL), and the cells were incubated for 20 min in the dark at room temperature. After PBS washing, in 10 randomly selected fields, the number of BrdU-positive cells and the total number of cells were counted under the microscope, and the percentage of BrdU-positive cells was calculated.

Li W., Hu X, & Huang X. (2022). Long intergenic non-protein coding RNA 847 promotes laryngeal squamous cell carcinoma progression through the microRNA-181a-5p/zinc finger E-box binding homeobox 2 axis. Bioengineered, 13(4), 9987-10000.

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Cell Proliferation Assay with BrdU Staining

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MDA-MB-231 and MCF-7 cells were inoculated into 24-well plates (with a cover glass inside) at 2.5 × 10⁵ cells/well. 24 h later, BrdU solution (Beyotime Biotechnology, Shanghai, China) was loaded, followed by incubation for 4 h. Subsequently, the cells were fixed with 4% paraformaldehyde for 30 min. Next, the cells were stained with 100 μL of 1 × Apollo for 30 min. Next, l g/mL DAPI staining solution was dripped into each well for incubation in darkness for 20 min at ambient temperature. The cells were washed with Phosphate Buffer Saline (PBS), and the total number of cells and the number of BrdU-positive cells in the randomly chosen 10 high-power fields were counted under a microscope. The positive rate of BrdU staining was then calculated.

Zhang H., Wang M., Lang Z., Liu H., Liu J, & Ma L. (2022). MiR-135a-5p suppresses breast cancer cell proliferation, migration, and invasion by regulating BAG3. Clinics, 77, 100115.

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Quantifying Cell Proliferation via BrdU

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The single-cell suspensions of A498 and ACHN cells were prepared and inoculated into a 96-well plate (1×10⁴ cells/well). Twenty-four hours after incubation, BrdU solution (30 μmol/L, Beyotime, Shanghai, China) was added into the cells for 8-hour incubation at 37° C. Next, the cells were fixed by 4% paraformaldehyde at indoor temperature for 30 min before they were flushed with PBS three times. 0.1% Tween-20 was utilized for permeation for 30 min at 37° C. After that, 5% BSA (Sigma-Aldrich) was applied to block the cells for 1 h at indoor temperature, which were then incubated along with the primary anti-BrdU antibody (1:500; cat. no. ab6326; Abcam) for 2 h at 37° C. After being rinsed three times with PBS, the cells were incubated with Alexa Fluor® 488 Rat monoclonal [BU1/75 (ICR1)] to BrdU (1:200, ab220074, Abcam) for 1 h at indoor temperature. DAPI staining solution (Beyotime, Shanghai, China) was harnessed to dye the nuclei at 37° C for 30 min. A fluorescence microscope (magnification, ×400; Olympus Corporation) was manipulated to observe the cells and capture their images. Cell proliferation rate = the number of BrdU staining positive cells/total DAPI staining positive cells ×100%.

Sun F., Sun P., Yang X., Hu L., Gao J, & Tian T. (2021). Inhibition of FSTL3 abates the proliferation and metastasis of renal cell carcinoma via the GSK-3β/β-catenin signaling pathway. Aging (Albany NY), 13(18), 22528-22543.

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Cell Proliferation and Migration Assays

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Cell proliferation was measured by the BrdU method. In brief, VSMCs were inoculated into a 35 mm culture dish containing a slide and then incubated with BrdU solution (Beyotime, Shanghai, China) for 4 h. Subsequently, cells were fixed in paraformaldehyde for 10 min and then incubated with the anti-BrdU antibody for 2 h at room temperature. Cells were then stained in DAPI buffer (Beyotime, Shanghai, China). Following several washes in PBS, five visions were selected randomly under the fluorescent microscope (Olympus, Tokyo, Japan) to calculate the average quantity of BrdU-positive cells.
Cell migration was evaluated by the Transwell system. Specifically, 600 μL RPMI-1640 medium containing 10% FBS was added into the lower chambers, while in the upper chambers, 100 μL serum-free RPMI-1640 medium containing 10 15 cells was added. Following 12 hours of culture at 37°C, cells that remained in the upper chambers were removed by a cotton swab, while those in the lower chambers were fixed in 4% paraformaldehyde for 10 min and stained in 0.5% crystal violet. Eventually, 5 visions were selected from each membrane randomly to calculate the average quantity of migrated cells.

Deng H., Xu H, & Luo Y. (2022). Mechanisms of miR-29a-5p involvement in osteogenic phenotype transformation and cellular regulation of vascular smooth muscle and thus influencing calcification in VSMCs in chronic kidney disease. Cellular and molecular biology (Noisy-le-Grand, France), 68(7).

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