Log In Sign up
The largest database of trusted experimental protocols

Hoechst

Manufactured by RiboBio
Visit Supplier
Sourced in China

Hoechst is a fluorescent dye used for nucleic acid staining in various laboratory techniques. It binds to the minor groove of DNA and emits fluorescence when excited, allowing the visualization and detection of DNA in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

5 ethynyl 2 deoxyuridine (edu), by RiboBio (3 mentions) Apollo dye solution, by RiboBio (2 mentions) Triton x 100, by Beyotime (2 mentions) Confocal microscope, by Leica (2 mentions) Imagequanttl7.0 image analysis software, by GE Healthcare (1 mentions) Cck 8 solution, by Beyotime (1 mentions) Triton x 100, by Solarbio (1 mentions) Fluorescence microscope, by Thermo Fisher Scientific (1 mentions) Click itr edu kit, by RiboBio (1 mentions) Inverted fluorescence microscope, by Zeiss (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by Zeiss (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions) Evos fl imaging system, by Thermo Fisher Scientific (1 mentions) Apollo 567 reaction cocktail, by RiboBio (1 mentions) Edu reagent, by RiboBio (1 mentions) Microplate reader, by Agilent Technologies (1 mentions) Cck 8, by Agilent Technologies (1 mentions) Cell light edu apollo567 in vitro kit, by RiboBio (1 mentions) Cck 8 kit, by Dojindo Laboratories (1 mentions) Fluorescence microscope, by Nikon (1 mentions)

Close spelling variants of this product

Hoechst 33342 Hoechst reagent Hoechst 33342 reaction solution

15 protocols using hoechst

1

Evaluating Cellular Proliferation and Clonogenicity

Check if the same lab product or an alternative is used in the 5 most similar protocols
For CCK8 assay, HCT-116 cells and HCT-116 corrected clone cells were seeded into 96-well cell culture plates (5000 cells/well). After 24, 48, and 72 hrs, CCK8 solution (Beyotime, China) was added for an additional 4 hrs at 37°C. Absorbance at 450 nm was determined for each well using a microplate enzyme-linked immunosorbent assay reader.
For EDU dyeing, HCT-116 cells and HCT-116 corrected clone cells were seeded into 96-well cell culture plates (5000 cells/well). After 24 hrs, 25 µM 5-ethynyl-2ʹ-deoxyuridine (EDU) was added for an additional 2 hrs at 37°C. Then, the cells stain with Cell-LightTM Apollo Stain Kit (RIBOBIO, Guangzhou, China) after washed with PBS and fixed with 4% formaldehyde for 30 mins. Before stained with Hoechst (RIBOBIO, Guangzhou, China) for 15mins, the cells were permeabilized with 0.5% Triton X-100 (Solarbio, Beijing, China) for 15 min. Images were obtained using a fluorescent microscopy (BX-51, Olympus Corporation, Tokyo, Japan).
For clonogenic assay, cells were plated onto 6-well plates (500 cells/well). Plated cells were cultured in 5% CO2 with 95% humidity at 37°C for 14 days. The plates were then fixed with 4% formaldehyde and stained with Giemsa. The colonies containing more than 50 cells were counted using ImageQuantTL7.0 Image Analysis Software (GE Healthcare, Buckinghamshire, UK).
Li Y., Li X., Qu J., Luo D, & Hu Z. (2020). Cas9 Mediated Correction of β-catenin Mutation and Restoring the Expression of Protein Phosphorylation in Colon Cancer HCT-116 Cells Decrease Cell Proliferation in vitro and Hamper Tumor Growth in Mice in vivo. OncoTargets and therapy, 13, 17-29.
+ Open protocol
+ Expand
2

Cell Proliferation Quantification using EdU Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The 5-ethynyl-2′-deoxyuridine (EdU) assay was performed using the Click-iTR EdU Kit (RiboBio, China) according to the manufacturer’s instructions. Specifically, cells were plated in 96-well plates and treated after adhesion. A total of 100 μL of culture medium containing 50 mM EdU was added to each well, and, 4 h later, the cells were fixed with 4% formaldehyde for 30 min. After washing, the cells were incubated with a solution in the kit for 30 min and stained with Hoechst (RiboBio, China) to identify nuclei, and images were captured with a fluorescence microscope (Thermo Fisher Scientific, USA). EdU-positive cells were determined using ImageJ 1.50i software (https://imagej.en.softonic.com/).
Hu M., Zheng Y., Liao J., Wen L., Cheng J., Huang J., Huang B., Lin L., Long Y., Wu Y., Ye X., Fu Y., Qi H., Baker P.N, & Tong C. (2022). miR21 modulates the Hippo signaling pathway via interference with PP2A Bβ to inhibit trophoblast invasion and cause preeclampsia. Molecular Therapy. Nucleic Acids, 30, 143-161.
+ Open protocol
+ Expand
3

EdU Proliferation Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells (5 × 104) were seeded in 24-well plates and cultured overnight. Then 300 µl EdU (50 µM)(RiboBio, China) was added in each well and incubated for 2 h. Then cells were fixed by 4% paraformaldehyde for 20 min and permeabilized by 0.5% TritonX-100 (Beyotime, China) for 20 min, dyed in 300 µl Apollo dye solution (RiboBio, China) for 25 min. Cell nuclei were dyed with Hoechst (RiboBio, China) for 10–30 min. Cells were observed under an inverted fluorescence microscope (Zeiss, Germany) at a magnification of 200×. And then proportion of EdU positive cells was calculated.
Wang H., Li H., Jiang Q., Dong X., Li S., Cheng S., Shi J., Liu L., Qian Z, & Dong J. (2021). HOTAIRM1 Promotes Malignant Progression of Transformed Fibroblasts in Glioma Stem-Like Cells Remodeled Microenvironment via Regulating miR-133b-3p/TGFβ Axis. Frontiers in Oncology, 11, 603128.
+ Open protocol
+ Expand
4

EdU Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded at a density of 2×105 cells/well in 6-well plates and incubated for 24 h. The cells were then treated with 50 µM EdU reagent (Guangzhou RiboBio Co., Ltd., Guangzhou, China) followed by Apollo-567 reaction cocktail (Guangzhou RiboBio Co., Ltd.) for 30 min. Following three washes with phosphate-buffered saline, cells were counterstained with Hoechst (dilution 1:1,000; Guangzhou RiboBio Co., Ltd.) for 10 min at room temperature for nuclear staining. All cells were observed using an EVOS FL Imaging System (Thermo Fisher Scientific, Inc.).
Lv Q., Wang G., Zhang Y., Han X., Li H., Le W., Zhang M., Ma C., Wang P, & Ding Q. (2019). FABP5 regulates the proliferation of clear cell renal cell carcinoma cells via the PI3K/AKT signaling pathway. International Journal of Oncology, 54(4), 1221-1232.
+ Open protocol
+ Expand
5

Proliferation Assays of VSMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell-Light EdU Apollo567 In Vitro Kit (RiboBio, Guangzhou, China) and CCK-8 kit (Dojindo, Shanghai, China) were used to detect the proliferation of VSMCs following the manufacturers’ protocols. Briefly, for EdU assay, proliferating cells were labeled with EdU, and Hoechst (RiboBio, Guangzhou, China) was used to label all cells. Five fields were randomly selected to take pictures under a fluorescence microscope (Nikon, Tokyo, Japan) and analyzed using ImageJ software. For CCK-8 assay, treated VSMCs were seeded in 96-well plates at 8 × 103 cells/well for 24 h and then incubated with CCK-8 solution at 37 °C for 2 h. Absorbance at 450 nm was measured using a microplate reader (Bio-Tek, CA, USA).
Rong Z., Li F., Zhang R., Niu S., Di X., Ni L, & Liu C. (2023). Ant-Neointimal Formation Effects of SLC6A6 in Preventing Vascular Smooth Muscle Cell Proliferation and Migration via Wnt/β-Catenin Signaling. International Journal of Molecular Sciences, 24(3), 3018.
+ Open protocol
+ Expand
6

Intracellular Localization of lncRNA

Check if the same lab product or an alternative is used in the 5 most similar protocols
Digoxin-labeled probe (Exon Biotechnology Inc., Guangzhou, China) was used to identify the intracellular localization of lncRNA. Cells were treated with 0.5% Triton X-100, 20 µg/ml proteinase K, and 4% PFA for 5 min, and then soaked with 3% H2O2 for 30 min. The following conditions were used: Probe: Hybridization solution=1:50 dilution, 5-min denaturation at 88°C, 10-min equilibration at 37°C, and 24-h hybridization. Cells were incubated with 3% BSA for 30 min at 37°C, followed by 1-h culture with Avidin-HRP: 1% BSA=1:100 dilution, 15-min culture with TSA (green): 0.15% H2O2: TSA dilution=1:1:100, and final counterstaining with Hoechst (Guangzhou RiboBio Co., Ltd.) for 15 min at room temperature. The cells were then observed under a confocal microscope (Leica Microsystems GmbH).
Zhou T., Zhang J., Qin B., Xu H., Zhang S, & Guan H. (2020). Long non-coding RNA NONHSAT143692.2 is involved in oxidative DNA damage repair in the lens by regulating the miR-4728-5p/OGG1 axis. International Journal of Molecular Medicine, 46(5), 1838-1848.
+ Open protocol
+ Expand
7

Visualizing Cell Proliferation with EdU

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells at 50% confluency were cultured with EdU labeling medium (RiboBio, Guangzhou, China) for 2 hours, fixed with 4% paraformaldehyde for 30 min and treated with 1% Triton X‐100 for 30 min at room temperature. After 3 washes with PBS, the cells were dyed with Apollo (RiboBio) for 30 min. Then, Hoechst (RiboBio) was used to stain the DNA in the cells, and subsequently visualized with a confocal microscope (Leica).
Liu C., Shen A., Song J., Cheng L., Zhang M., Wang Y, & Liu X. (2023). LncRNA‐CCAT5‐mediated crosstalk between Wnt/β‐Catenin and STAT3 signaling suggests novel therapeutic approaches for metastatic gastric cancer with high Wnt activity. Cancer Communications, 44(1), 76-100.
+ Open protocol
+ Expand
8

Quantifying Cell Proliferation by EdU Staining

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seed 200 μl in a 48‐well plate at a density of 1.5×105/ml. The cells were incubated 2 h by EDU marketing solution (Ribobio, #C10310‐1), and then fixed by 4% paraformaldehyde for 30 min, and the stain solution was added in the plate for 30 min. Following three washes with PBS, cells were counterstained with Hoechst (dilution 1:1000; Ribobio, # C10310‐1) for 10 min at room temperature for nuclear staining. Then observe the photo under a fluorescence microscope (Olympus, IX73).
Gui D., Dong Z., Peng W., Jiang W., Huang G., Liu G., Ye Z., Wang Y., Xu Z., Fu J., Luo S, & Zhao Y. (2021). Ubiquitin‐specific peptidase 53 inhibits the occurrence and development of clear cell renal cell carcinoma through NF‐κB pathway inactivation. Cancer Medicine, 10(11), 3674-3688.
+ Open protocol
+ Expand
9

Measuring Cell Proliferation via EdU Incorporation

Check if the same lab product or an alternative is used in the 5 most similar protocols
To assess cell proliferation, MCF-7 cells were seeded into 48-well plates. The cells were incubated under standard conditions in complete media (DMEM supplemented with 10% FBS). Transfection of the cells was performed the following day as described above. Twenty-four hours after transfection, the cells transfected with miRNA were harvested. Forty-eight hours after transfection, the cells transfected with siRNA and overexpression plasmid were harvested. Cell proliferation was detected by the incorporation of 5-ethynyl-2′-deoxyuridine (EdU) using the EdU Cell Proliferation Assay Kit (Ribobio, Guangzhou, China). Briefly, the cells were incubated with 50 μM EdU for 8 h before fixation, permeabilization and EdU staining, which were performed according to the manufacturer’s protocol. The cell nuclei were stained with Hoechst (Ribobio, Guangzhou, China). The proportion of nucleated cells incorporating EdU was determined by fluorescence microscopy.
Hong Y., Liang H., U.z.a.i.r.-.u.r.-.R.e.h.m.a.n., Wang Y., Zhang W., Zhou Y., Chen S., Yu M., Cui S., Liu M., Wang N., Ye C., Zhao C., Liu Y., Fan Q., Zhang C.Y., Sang J., Zen K, & Chen X. (2016). miR-96 promotes cell proliferation, migration and invasion by targeting PTPN9 in breast cancer. Scientific Reports, 6, 37421.
+ Open protocol
+ Expand
10

Cell Proliferation Assay with EdU

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded into 24-well plates at a density of 5×104 cells/well and incubated overnight, after which 300 μl of EdU (50 μM) (RiboBio, China) were added to each well, and the cells were incubated for additional 2 h. The cells were then fixed in 4% polyformaldehyde for 20 min, permeabilized with 0.5% TritonX-100 (Beyotime, China) for 20 min, and stained in 300 μl of Apollo dye solution (RiboBio, China) for 25 min. Cell nuclei were stained with Hoechst (RiboBio, China) for 10-30 min. The proportion of EdU-positive cells were determined using a fluorescence microscope.
Wang H., Tan L., Dong X., Liu L., Jiang Q., Li H., Shi J., Yang X., Dai X., Qian Z, & Dong J. (2020). MiR-146b-5p suppresses the malignancy of GSC/MSC fusion cells by targeting SMARCA5. Aging (Albany NY), 12(13), 13647-13667.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!