Product Description
The upgraded Cell Cycle Assay Kit Plus uses RedNucleus I staining method to detect cell cycle. RedNucleus I is a far-infrared nucleic acid dye with cell membrane permeability, which can quickly enter live cells, specifically bind to DNA, and perform cycle detection on live cells without the need for RNase digestion. Compared with the traditional PI staining method, cells do not require membrane rupture or fixation, making the operation more convenient.
RedNucleus I is a fluorescent dye for double stranded DNA, and the fluorescence intensity after binding with double stranded DNA is proportional to the content of double stranded DNA. The intracellular DNA content can be measured by flow cytometry, and then cell cycle analysis can be performed based on the distribution of DNA content. After RedNucleus I staining, assuming that the fluorescence intensity of G0/G1 phase cells is 1, the theoretical value of fluorescence intensity for G2/M phase cells containing double genomic DNA is 2, and the fluorescence intensity for S phase cells undergoing DNA replication is between 1-2. In addition, RedNucleus I is compatible with dyes such as Horizon BV/BUV, FITC, and R-PE, and can undergo periodic testing after sample staining.
This kit is usually used for cell cycle detection of cultured adherent or suspended cells. If used for cell cycle detection of tissues, the tissues must first be digested into a single-cell state.
ordering information
Product Name | Article number | Specifications |
Cell cycle detection kit Plus (Cell Cycle and Apoptosis Kit Plus) | AC12L553 | 50 T |
Product components
Composition | Specifications |
A. Staining buffer (10 x) | 10 mL |
B. RedNucleus I staining solution | 200 μL |
Transportation and Preservation
Blue ice transportation- Store at 20 ℃, component B needs to be stored away from light. The validity period is 24 months. [Note]: Please store at 4 ℃ for short-term use.
usage method
1. Preparation of cell samples:
(1) (This step is for adherent cells. If it is a suspended cell, the second step can be performed directly.) Digest the cells with trypsin, add cell culture medium, gently disperse the cells, and collect them in a centrifuge tube.
(2) Centrifuge around 1000 g for 3-5 minutes to precipitate cells. Carefully remove the supernatant, add approximately 1 mL of pre cooled 1 x staining buffer in an ice bath (dilute 10 x staining buffer with diH2O at a ratio of 1:10), and resuspend the cells. Repeat once.
(3) Centrifuge around 1000 g for 3-5 minutes to precipitate cells. After discarding the supernatant, add 1 mL of culture medium to resuspend the cells (for fixed cells, 1 x PBS can also be used for resuspend). Gently tap the bottom of the centrifuge tube to disperse cells appropriately and avoid cell clumping.
2. Staining: Add 4 μ L of RedNucleus I staining solution to each cell sample, mix slowly and thoroughly, and incubate at room temperature in the dark for 20 minutes (or at 37 ℃ in the dark for 5-10 minutes).
3. Flow cytometry detection and analysis: Use a flow cytometer to excite at 638 nm. It is recommended to detect in RL3 or FL4 channels, and RL1 and RL2 channels can also be used for detection. Use appropriate analysis software for cell DNA content analysis and light scattering analysis.
matters needing attention
1. This product is limited to scientific experimental research and cannot be used in clinical diagnosis, treatment, or other fields.
2. This product is suitable for detecting both live and fixed cell cycles. If fixation is required, it is recommended to use ice bath pre cooling with 75-80% ethanol overnight to fix the cells.
3. Fluorescent dyes all have quenching problems. Please try to avoid light during storage and use to slow down fluorescence quenching.
This product is limited to scientific experimental research and cannot be used in clinical diagnosis, treatment, or other fields.
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