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BODIPY 505/515

CAS No. : 21658-70-8

MCE 国际站：BODIPY 505/515

产品活性：BODIPY505/515 是一种 BODIPY 染料。BODIPY 染料是一种具有强紫外吸收能力的小分子染料，其荧光峰比较尖锐，量子产率高。其对环境的极性和 pH 值相对不敏感，因此，在不同生理条件下相对稳定。由于其结构的非对称性，BODIPY 具有多种衍生结构产物。BODIPY 脂滴类染料可以很好的穿过细胞膜进入细胞内部，通过在细胞内的极性脂类上定位以进行特异性染色，因此，该染料可用于标记活细胞和固定细胞。最大激发/发射波长：505/515 nm。

研究领域：Others

作用靶点：Fluorescent Dye

In Vitro: General Protocol
1. Preparation of BODIPY 505/515 working solution
1.1 Preparation of the stock solution
Dissolve 1 mg BODIPY 505/515 in 382 μL DMSO to obtain 10 mM of stock solution.
Note: It is recommended to store the stock solution at -20°C or -80°C, keep away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of BODIPY 505/515 working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of working solution.
Note: Please adjust the concentration of BODIPY 505/515 working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells (6-well plate)
a. Centrifuge at 1000 g at 4°C for 3-5 min and then discard the supernatant. Wash twice with PBS, 5 min each time.The cell density is 1×10⁶/mL.
b. Add 1 mL of working solution, and then incubate at room temperature for 5-30 min.
c.Centrifuge at 400 g at 4°C for 3-4 min and then discard the supernatant.
d. Wash twice with PBS, 5 min each time.
e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 5-30 min.
d. Wash twice with medium, 5 min each time. Observation by fluorescence microscopy or flow cytometry.
2.3 TissueSlice（Paraffin sections）
2.3.1 Room temperature dewaxing and hydration: Xylene I for 10 min, Xylene II for 8 min, Xylene III for 8 min, Anhydrous Ethanol for 5 min, 90% Ethanol for 3 min, 80% Ethanol for 3 min, 70% Ethanol for 3 min. Distilled water 3 min*3 times,PBS 3 min*3 times.
2.3.2 For thermal antigen repair, replace the section holder with a new one and put in the hydrated sections. Add antigen repair solution and heat and boil. Control the temperature at 96-98℃. The sections are then placed in the hot pot for 15 min and finally cooled naturally at room temperature. five min in PBS*2 times and 3 min in distilled water*2 times.
2.3.3 Immunohistochemistry pen circle: Remove the tissue, shake off the water, and use absorbent paper to absorb the water droplets. Draw circles around the tissue with the histochemistry pen, with the circles intact.
2.3.4 Inactivation of endogenous enzyme activity: Place the sections in a wet box with a small amount of distilled water, add 3% hydrogen peroxide dropwise and incubate for 10 min at room temperature. three min*3 times with PBS and three min with distilled water.
2.3.5 Add 50-100 μM dye to the tissue and incubate for 30-60 min.
2.3.6 Aspirate the dye working solution and wash 2-3 times with PBS or culture medium for 5 min each time.
2.3.7 Perform imaging observations using fluorescence microscopy.
Storage
-20°C, 1 year; Protect from light.
Precautions
1. Please adjust the concentration of BODIPY 505/515 working solution according to the actual situation.
2. Experiments suggest a positive control, incubate the control group cells with 30 μM oleic acid for 8 h and then perform subsequent experiments.
3. This product is only for R&D use, not for drug, household, or others.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.

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