基本描述

规格或纯度

BioReagent, 用于显微镜, 生物染色剂, 适用于免疫荧光(IF), 适用于荧光分析, 用于细胞培养, 即用型, for DNA and RNA applications, 无菌过滤

稳定性与储存

Store at -20℃ long term (12 months). Store in the dark. Avoid repeated freezing and thawing.

英文名称

Cell Cycle and Apoptosis Analysis Kit

储存温度

避光,-20°C储存,避免反复冻融

运输条件

超低温冰袋运输

产品介绍

细胞周期与细胞凋亡检测试剂盒（Cell Cycle and Apoptosis Kit）是一种采用经典的碘化丙啶（PropidiumIodide，PI）染色方法进行细胞周期与细胞凋亡分析的检测试剂盒。碘化丙啶是一种双链DNA的荧光染料。碘化丙啶和双链DNA结合后可以产生荧光并且荧光强度和双链DNA的含量成正比。细胞内的DNA被碘化丙啶染色后，可以用流式细胞仪对细胞进行DNA含量测定，然后根据DNA含量的分布情况，可以进行细胞周期和细胞凋亡分析。碘化丙啶染色后，假设G0/G1期细胞的荧光强度为1，那么含有双份基因组DNA的 G2/M期细胞的荧光强度的理论值为2，正在进行DNA复制的S期细胞的荧光强度为1~2 之间。凋亡细胞由于细胞核发生浓缩以及发生DNA片段化（DNAfragmentation）导致部分基因组DNA片段在染色过程中丢失，因此凋亡细胞碘化丙啶染色后呈现明显的弱染，即荧光强度小于1，在流式检测的荧光图上出现所谓的sub-G1峰，即凋亡细胞峰。细胞发生凋亡时，由于胞浆和染色质浓缩、核碎裂，产生凋亡小体，使细胞的光散射性质发生变化。在细胞凋亡的早期，细胞对前向角光散射的能力显著降低，对侧向光散射的能力增加或没有变化。在细胞凋亡的晚期，前向和侧向光散射的信号均降低，因此可通过流式细胞仪测定细胞光散射的变化观察细胞凋亡情况。本试剂盒通常应用于培养的贴壁或悬浮细胞的细胞周期与细胞凋亡检测。如果用于组织的细胞周期与细胞凋亡检测，则必须把组织消化成单细胞状态，才可以进行检测。

P1373478	组分	20 T	50 T	100 T	Storage	Quantity Per Test
P1373478A	染色缓冲液	10 mL	25 mL	50 mL	-20℃.	500 μL per 1 × 10⁶ cells.
P1373478B	碘化丙啶染色液(20X)	100 μL	250 μL	500 μL	-20℃.避光保存.	5 μL per 1 × 10⁶ cells.
P1373478C	RNase A (50×)	0.2 mL	0.5 mL	1 mL	-20℃.	10 μL per 1 × 10⁶ cells.

注: 每个Test推荐染色的细胞数量为 1 × 10⁶ cells.

使用方法

1. 收集细胞，预冷PBS清洗两次；按照0.1–1.0 × 10⁷的密度用预冷PBS重悬；

2. 在细胞悬液中，边震荡边逐滴加入无水乙醇，使乙醇最终浓度为70%；

3. 轻柔混匀细胞，4℃过夜；

4. 预冷PBS清洗细胞两次后，按照2 × 10⁶/mL的密度用染色缓冲液重悬；

5. 每500 μL 细胞悬液中加入5 μL 碘化丙啶染色液及10 μL RNase A；

6. 缓慢并充分重悬细胞，室温避光孵育30min；

7.流式检测和分析。

注：需自备PBS和无水乙醇。

注意事项

1. 荧光染料均存在淬灭问题，保存和使用过程中请尽量注意避光，以减缓荧光淬灭。

2. 碘化丙啶对人体有刺激性，请注意适当防护。

3. 碘化丙啶是已知的诱变剂，因此该溶液在丢弃之前需要先经过活性炭处理。

4. 70%乙醇固定细胞一定要充分，否则染色不均导致结果不明显或者偏差。建议 70%乙醇-4℃固定过夜。

5. 细胞固定后要保证细胞是单细胞悬液，细胞的粘连可能会影响我们的结果。

6. 为了您的安全和健康，请穿实验服并戴一次性手套操作。

7. 本产品仅限于科研，不得用于临床诊断或治疗，不得用于食品和药品，不得存放于普通住宅内。

8. 为了您的安全和健康，请穿实验服并戴一次性手套操作

The Cell Cycle and Apoptosis Kit (Cell Cycle and Apoptosis Kit) is a test kit that uses the classic propidium iodide (PI) staining method to analyze the cell cycle and apoptosis. Propidium iodide is a fluorescent dye for double-stranded DNA. When propidium iodide binds to double-stranded DNA, it can produce fluorescence, and the fluorescence intensity is proportional to the content of double-stranded DNA. After the DNA in the cells is stained with propidium iodide, the DNA content of the cells can be measured using a flow cytometer, and then based on the distribution of DNA content, the cell cycle and apoptosis can be analyzed. After staining with propidium iodide, if the fluorescence intensity of G0/G1 phase cells is 1, then the theoretical fluorescence intensity of G2/M phase cells containing double genomic DNA is 2, and the fluorescence intensity of S phase cells undergoing DNA replication is between 1 and 2. Apoptotic cells, due to the condensation of the cell nucleus and the fragmentation of DNA (DNA fragmentation), cause some genomic DNA fragments to be lost during staining, so the iodine staining of apoptotic cells presents a significantly weak color, that is, the fluorescence intensity is less than 1, and a so-called sub-G1 peak appears on the fluorescence graph of flow cytometry, which is the peak of apoptotic cells. When cells undergo apoptosis, due to the concentration of cytoplasm and chromatin, apoptotic bodies are produced, causing changes in the light scattering properties of the cells. In the early stage of cell apoptosis, the ability of the cell to forward-angle light scattering significantly decreases, while the ability of lateral light scattering increases or remains unchanged. In the late stage of cell apoptosis, the signals of forward and lateral light scattering both decrease, so the changes in cell light scattering can be measured using a flow cytometer to observe the apoptosis situation. This kit is usually applied to the detection of the cell cycle and apoptosis of cultured adherent or suspended cells. If used for the cell cycle and apoptosis detection of cells from tissues, the tissue must be digested into a single-cell state before detection can be performed.

P1373478	Component	20 T	50 T	100 T	Storage	Quantity Per Test
P1373478A	Dyeing buffer solution	10 mL	25 mL	50 mL	-20℃.	500 μL per 1 × 10⁶ cells.
P1373478B	Propidium Iodide staining solution (20X)	100 μL	250 μL	500 μL	-20℃.Store in the dark.	5 μL per 1 × 10⁶ cells.
P1373478C	RNase A (50X)	0.2 mL	0.5 mL	1 mL	-20℃.	10 μL per 1 × 10⁶ cells.

Note: The recommended number of cells to stain per test is 1 × 10⁶ cells

Instructions for use

1.Collect cells and wash twice with ice-cold PBS; resuspend in ice-cold PBS at a density of 0.1–1.0 × 10⁷ cells/mL.

2.In the cell suspension, anhydrous ethanol was added dropwise while shaking until the final ethanol concentration reached 70%.

3.Mix gently and fix overnight at 4 °C.

4.Wash cells twice with ice-cold PBS, then resuspend in dyeing buffer solution of 2 × 10⁶ cells/mL.

5.Add 5 µL of Propidium iodide and 10 µL RNase A to 500 µL of the cell suspension.

6.Resuspend cells gently and thoroughly, then incubate at room temperature for 30 min in the dark.

7.Analyze samples by flow cytometry.

Note: Please bring your own PBS and absolute ethanol.

Precautions

1. Fluorescent dyes all have the problem of quenching. Please try to avoid light during storage and use to slow down fluorescence quenching.

2. Iodophor is irritating to the human body. Please take appropriate protective measures.

3. Iodophor is a known mutagen, so this solution needs to be treated with activated carbon before being discarded.

4. When fixing cells with 70% ethanol, make sure it is thorough; otherwise, uneven staining will result in unclear or inaccurate results. It is recommended to fix the cells overnight at -4℃ with 70% ethanol.

5. After cell fixation, ensure that the cells are in a single-cell suspension; cell adhesion may affect our results.

6. For your safety and health, please wear laboratory coats and disposable gloves during operation.

7. This product is only for scientific research and cannot be used for clinical diagnosis or treatment, nor for food or medicine. It must not be stored in ordinary residences.

8. For your safety and health, please wear laboratory coats and disposable gloves during operation.

图片

Flow Cytometry: Cell Cycle and Apoptosis Detection Kit (P1373478)

Cell Cycle and Apoptosis Detection Kit (P1373478) - Flow Cytometry
Jurkat cells were stained with the “Cell Cycle and Apoptosis Detection Kit” (P1373478). The resulting graph depicts the DNA content distribution of the cell population. Following overnight fixation in 70% ethanol, the cells were resuspended in a staining buffer containing Propidium Iodide (PI) and RNase A, and then analyzed.
In the graph, the horizontal axis represents DNA content (measured by PI fluorescence intensity), and the vertical axis corresponds to the number of cells. The distinct peaks and regions correspond to different phases of the cell cycle:
1. Sub-G1 phase: Apoptotic cells with fractional DNA content
2. G0/G1 phase: This peak corresponds to quiescent state or gap 1 (G1) phases, containing a diploid (2N) DNA content.
3. S phase: This region between the G0/G1 and G2/M peaks represents cells actively synthesizing DNA. Their DNA content varies between 2N and 4N.
4. G2/M phase: This peak corresponds to cells in the gap 2 (G2) and mitotic (M) phases. These cells have completed DNA replication and contain a tetraploid (4N) DNA content.

名称和识别符

分子类型	生物试剂/缓冲液

化学和物理性质

敏感性	light-sensitive

质检证书（CoA,COO,BSE/TSE 和分析图谱)

C of A & Other Certificates(BSE/TSE, COO):

输入批号以搜索分析图谱:

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

=

浓度

x

体积

x

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

x

体积 1 (V1)

=

浓度 2 (C2)

x

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

=

质量（小瓶）

÷

所需的复溶浓度

货号 (SKU)	包装规格	是否现货
P1373478-20T	20T	期货
P1373478-50T	50T	期货
P1373478-100T	100T	期货

细胞周期与细胞凋亡检测试剂盒

库存信息

库存信息

库存信息

基本描述

图片

名称和识别符

化学和物理性质

质检证书（CoA,COO,BSE/TSE 和分析图谱)

溶液计算器

摩尔浓度计算器

稀释计算器

复溶计算器