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文献和实验Purpose Materials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS 43 ml blood 12 ml RT Histopaque 107718 ml cold H2 O2 ml 10x PBSM199 for HUVECs: 1L powder pocket
To fix, 1/10 volume of 37 formaldehyde is added to the culture, which is shaken for a further 40 minutes at room temperature. Cells are then recovered by centrifugation at 1400 X g for 2 minutes, and washed (i.e. gently resuspended then recovered
1.5M Tris(PH 8.8) 100ml 18.171g (at4°C) 10% SDS 100 ml 10g (at room temperature) 10% ammonium persulfate 1ml 0.1g(replace every week store at at4°C) TEMED 8 room temprature Antibodies Primary antibody: 5A11(anti-mouse IgG) 500x-1000x Secondary
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