BioReagent,即用型,生物染料级,生物染色剂,用于显微镜,用于细胞培养,适用于免疫荧光(IF),适用于荧光分析 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 33 篇同行评审文献引用。
Hoechst 33258是一种蓝色至蓝绿色荧光的活细胞染料，可标记DNA。Hoechst 33258可特异性结合DNA小沟(同时倾向于结合富含A/T的DNA)，使荧光强度显著增强。Hoechst 33258可穿过细胞膜，导致DNA结构变化，如G2/M期阻滞。Hoechst 33258可结合活细胞或固定细胞，荧光强度随着溶液pH升高而增强。Hoechst 33258作为DNA特异性探针，可用于检测DNA含量、分析细胞周期等。Hoechst 33258的激发波长为350-365 nm，发射波长为460-490 nm。
本产品浓度为1.0 mg/mL，用于细胞核染色时，推荐工作浓度为1-10 μg/mL。
使用方法
1.对于固定的细胞或组织
a. 将细胞接种于培养板/皿中，培养至对数生长期。如需固定细胞，用4%多聚甲醛室温固定10-15 min，PBS洗涤2次，每次5分钟。
b. 加入Hoechst 33258工作液，37°C避光孵育10-30 min。
c. 吸除Hoechst 33258染色液，用PBS洗涤2-3次，每次5 min。
2.对于活细胞或组织
a. 收集细胞，1000 rpm离心5 min，弃上清。
b. 用PBS重悬细胞，加入Hoechst 33258工作液，室温避光孵育15-20 min。
c. 1000 rpm离心5 min，弃染色液，用PBS洗涤2-3次；加入PBS重悬细胞。
d. 直接在荧光显微镜下观察或封片后荧光显微镜下观察。细胞发生凋亡时，会看到凋亡细胞的细胞核呈致密浓染，或呈碎块状致密浓染。
注意事项
1.根据Hoechst 33258染色液应用的不同，建议使用前进行滴定以确定最佳工作浓度。
2.荧光染料都存在淬灭的问题，建议染色后尽量当天完成检测。活细胞或组织染色后宜立即观察。
3.本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
4.为了您的安全和健康，请穿实验服并戴一次性手套操作。
Hoechst 33258 is a blue to blue-green fluorescent live cell dye that can label DNA. Hoechst 33258 can specifically bind to the DNA groove (tending to bind to DNA rich in A/T at the same time), resulting in a significant increase in fluorescence intensity. Hoechst 33258 can pass through the cell membrane, causing changes in DNA structure, such as G2/M phase arrest. Hoechst 33258 can bind to live cells or fixed cells, and the fluorescence intensity increases with the increase of solution pH. As a DNA-specific probe, Hoechst 33258 can be used for detecting DNA content and analyzing cell cycle, etc. The excitation wavelength of Hoechst 33258 is 350-365 nm, and the emission wavelength is 460-490 nm.
The concentration of this product is 1.0 mg/mL. When used for nuclear staining, the recommended working concentration is 1-10 μg/mL.
Instruction for use
1.For fixed cells or tissues:
a. The cells were inoculated onto the culture plates/flasks and cultured until they reached the logarithmic growth phase. If cell fixation was required, the cells were fixed with 4% paraformaldehyde at room temperature for 10-15 minutes, then washed twice with PBS, each time for 5 minutes.
b. Add the Hoechst 33258 working solution and incubate at 37°C in the dark for 10-30 minutes.
c. Remove the Hoechst 33258 staining solution and wash the cells twice with PBS for 5 minutes each time.
2.For living cells or tissues：
a. Collect the cells, centrifuge at 1000 rpm for 5 minutes, and discard the supernatant.
b. Resuspend the cells in PBS, add the Hoechst 33258 working solution, and incubate at room temperature in the dark for 15-20 minutes.
c. Centrifuge at 1000 rpm for 5 minutes, discard the staining solution, wash twice with PBS for 5 minutes each time; then resuspend the cells in PBS.
d.Observe directly under a fluorescence microscope or under a fluorescence microscope after mounting. When cells undergo apoptosis, the nuclei of apoptotic cells will show dense and compact staining, or fragmented and dense staining.
Matters needing attention
1.Depending on the application of Hoechst 33258 staining solution, it is recommended to perform a titration before use to determine the optimal working concentration.
2.Fluorescent dyes all have the problem of quenching. It is recommended to complete the detection as soon as possible after staining. Live cells or tissues should be observed immediately after staining.
3.This product is only for professional scientific research use. It shall not be used for clinical diagnosis or treatment, nor for food or medicine. It shall not be stored in ordinary residences.
4.For your safety and health, please wear laboratory coats and disposable gloves during operation.
1-10 μg/ml
H302: 吞食有害
H315: 引起皮肤刺激
H319: 引起严重眼睛刺激
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ25F1129346 |
分析证书 | H1373484 |
| ZJ26F0737563 |
分析证书 | H1373484 |
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