Fluoro AChE檢測試劑盒

產品描述：

保存溫度：2-8度

主要優點：

• 無需放射性物質。
• 應用 - 流式細胞儀或熒光顯微鏡。
• 檢測細胞水平的細胞溶解活性。
• 適用于多種類型的哺乳動物細胞系。

技術

測量CMC / ADCCzui常用的方法是放射性鉻-51（51Cr）釋放試驗（2）。該測定有幾個缺點：昂貴的，難以加載某些細胞類型，由于嚴格的環境規定而處置昂貴，并且具有51Cr自發釋放的高背景。使用流式細胞儀，現在可以消除對放射性物質的需要，并增加在單個細胞基底上定量細胞溶解活性的能力。各組已經證明通過流式細胞術測量CMC / ADCC活性與傳統的51Cr釋放測定（3,4,5,6）具有強烈（95％）的相關性。

測定原理

細胞跟蹤染料CFSE（7,8,9）用于標記靶細胞群體。在測定完成實驗方案后，加入7AAD（活/死）（10,11）以測量細胞死亡。7AAD僅進入膜受損細胞并與DNA結合。

流式細胞術用于門靶細胞并測量7AAD陰性對7AAD后細胞。％細胞毒性通過以下等式計算（參見下面的實驗例）：

7AAD正（右上象限）= R1 / 7AAD Postitve（右上象限）= R1 + 7AAD負（右下象限）= R2 ×100 
ACT1

為了測試豬gd淋巴細胞的自然殺傷能力，將K562細胞染色并調整至含有10％FBS的1×10 4個細胞/100μlPRMI的終濃度。以25:1,50:1和100:1的E / T比加入gd淋巴細胞，調節至400μlRPMI的總體積，然后在37℃，無菌封蓋的面管中孵育4小時。孵育后，將活/死染色劑直接加入每個管中，在冰上孵育15分鐘并通過流式細胞術分析。

引用文獻

• a. Perussia, B., (1998). Current Topics in Microbiology and Immunology 230, p63.
  b. Whiteside, T.L., Rinaldo, C.R. and Herberman, R.B. (1992) Cytolytic Cellfunctions. In: N.R. Rose, E.C. de Macario (Eds.), Manual of Clinical Laboratory Immunology. American Society for Microbiology. Washington, DC, p. 220.
• Brunner, K.T., Manuel, J., Cerotini, J.C., Chapuis, B., (1968). Quantitative Assay of the Lytic Action of Immune Lymphoid Cells on Cr-labelled Allogenic Target Cells in- vitro; Inhibition by Iso-antibody and by Drugs, Immunology 14,181.
• Lee-MacAry, A.E., Ross, E.L, Davies, D., and Wilkinson, R.W., (2001). Development of a Novel Flow Cytometric Cell-mediated Cytotoxcity Assay Using the Fluorophores PKH-26 and TO-PRO-3 Iodide. J. Immunology. Met 252, 83-92.
• Gogoy-Ramirez, K., Franck, K., and Gains, H., (2000). A Novel Method for the Simultaneous Assessment of Natural Killer Cell Conjugate Formation and Cytotoxicity at the Single-cell Level by Multi-parameter Flow Cytometry. Journal of Immunology. Met 239, 35-44.
• Goldberg, J.E., Sherwood, S.W., Clayberger, C., (1999). A Novel Method for Measuring CTL and NK Cell-mediated Cytotoxicity Using Annexin V and Two-color Flow Cytometry. Journal of Immunology. Methods 224, 1.
• Hatam, L,. Schuval, S., Bonagura, V.R., (1994). Flow Cytometric Analysis of Natural Killer Cell Function as a Clinical Assay. Cytometry 16,59.
• L.S De Clerck et al.,J. Immunol. Meth. 172, 115 (1994).
• M. Bronner-Fraser,J. Cell Biol. 101, 610 (1985).
• Rabinovitch, P.S., et al., J. Immunol. 136, 2769 (1986).
• Su, X.,J. Immunol. 156, 156, 4198 (1996).
• Olin, MR. Lee, J. Choi, K, and Molitor, Tw. gd T-lymphocyte Cytotoxic Activity against Mycobaterium bovis Analysed by Flow Cytometry: Journal of Immunological Methods; Publication in process.
• Olin, MR. Thesis, K. Cho, J. and Molitor, T.W. Morphine Suppresses Microglial Directed Cytolytic Activity by gd Lymphocytes. Journal of Neuroimmunology; Publication in process.
•