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自旋捕獲劑

BMPO (Spin trapping reagent) 

產品簡介：

BMPO（5-tert-butoxycarbonyl 5-methyl-1-pyrroline N-oxide）是一種新型高效且高穩定型的硝酮自旋捕獲劑（Nitrone Spin Trapping Reagent），由美國威斯康星醫學院Kalyanaraman教授的實驗室團隊開發，非常適合用于特異性檢測和鑒定體內外形成的硫自由基、羥基自由基（?OH）和超氧陰離子自由基（?O₂-），通過形成用電子順磁共振波譜（EPR）可區分的加合物來測定。

BMPO具有以下特點：

①特異性高，半衰期長：其它的硝酮自旋捕獲劑比如：DMPO，難以輕易區分超氧陰離子和羥基自由基，因產生的DMPO-超氧加合物（半衰期t?=45s）瞬間衰減產生DMPO-羥基加合物。與最近開發的自旋捕獲劑：DEPMPO和EMPO類似，BMPO-超氧加合物不會衰減生成羥基加合物，但是，BMPO-超氧加合物的半衰期最長（t?=23min）。

②產品穩定性高：DEPMPO和EMPO是液體自旋捕獲劑，通常被硝基氧雜質污染，且效期有限。BMPO是固體的環形硝酮，通過結晶以高度純化的狀態提供，能夠保存更長的周期，且不用擔心降解。

③更高的信噪比：BMPO衍生的加合物在各自的EPR光譜中具有更高的信噪比，使其更適合用于檢測細胞懸浮液中的亞硫酸、羥基和甲基自由基。

④高水溶性：水溶性好，更利于水相體系自由基的研究，尤其是生物體系的自由基研究。

產品特性：

1） CAS NO：387334-31-8

2） 化學名：3,4-dihydro-2-methyl-1,1-dimethylethyl ester-2H-pyrrole-2-carboxylic acid-1-oxide

3） 同義名：5-tert-butoxycarbonyl 5-methyl-1-pyrroline N-oxide; BocMPO;

4） 分子式：C₁₀H₁₇NO₃

5） 分子量：199.2

6） 純度：≥98%

7） 外觀：結晶或結晶性粉末

8） 溶解性：溶于水、PBS（pH 7.2, 10mg/ml）、DMSO（25mg/ml）、無水乙醇（25mg/ml）

9） 化學結構圖：

保存條件: 

-20oC干燥保存，2年有效。 

注意事項：

1.      為了您的安全和健康，請穿實驗服并戴一次性手套操作。

應用示例（僅作參考）：

文獻1：

①黃嘌呤-黃嘌呤氧化酶體系內超氧陰離子（O2??）生成的測定：Typically, xanthine oxidase (0.04 U/mL) was added to pH ～7.4 sodium phosphate buffer (50 mM) containing DTPA (1 mM) and hypoxanthine (0.5–400 μM, final concentration) to achieve rates of O2?? formation that ranged from 0.1 to 6.0 μM/min. We estimated the superoxide formation rate by monitoring the SOD-inhibitable reduction of ferricytochrome c (80 μM) at room temperature. Spin trapping was performed by addition of 100 mM BMPO in pH ～7.4 phosphate-buffered saline (PBS; 50 mM) containing 1 mM DTPA to the solution of hypoxanthine and xanthine oxidase to achieve a final BMPO concentration of 50 mM in the reaction mixture. EPR spectra were recorded 10 min after mixing reagents. The half-life of BMPO-OOH at ambient temperature is reported to be ～23 min. Solutions for control experiments contained SOD (30 U/mL).

Figure 2 Comparison of CW and rapid-scan spectra of BMPO-OOH in solution with a O2?? production rate of 0.1μM/min, recorded 10 min after mixing reagents. The O2?? was produced by a hypoxanthine/xanthine oxidase mixture. The concentration of BMPO-OOH is ～0.3μM. (A) CW spectrum obtained with 55 G sweep width, 0.75 G modulation amplitude, single 30 s scan, 15 ms conversion time, 10 ms time constant, and 20 mW (B1 = 170 mG) microwave power. (B) Deconvolved rapid-scan spectrum obtained with 55 G scan width, 51 kHz scan frequency, and 53 mW (B1 = 250 mG) microwave power. Segments consisting of 12 sinusoidal cycles were averaged 100 k times, with a total data acquisition time of ～30 s.

文獻2：

①缺氧下的ESR測定（O2??，?OH）：For O2?? detection, the pre-deoxidized PBS (pH 5.0, 25?mM) contained 25?mM BMPO, 20?μg?mL?1 MnO2@PtCo nanoflowers, 100?μM H2O2, 50% DMSO was prepared. After incubation of 5?min, ESR spectra were recorded. For ?OH detection, the pre-deoxidized PBS (pH 5.0, 25?mM) contained 25?mM BMPO, 20?μg?mL?1 MnO2@PtCo nanoflowers, 100?μM H2O2, 0.25?U?mL?1 SOD was prepared. After incubation of 5?min, ESR spectra were recorded. The following instrument settings were used for collecting ESR spectra: 1?G field modulation, 100?G scan range, and 20?mW microwave power.

Fig f ESR spectra of BMPO/?OOH adducts from different groups in the hypoxic H2O2 (100?μM) condition upon addition of DMSO.g ESR spectra of BMPO/?OH adducts were collected from different samples in the hypoxic H2O2 (100?μM) condition upon addition of SOD. Data were presented as mean?±?s.d. (n=?5).