蛋白活性实验-TNF-a活性蛋白
蛋白活性实验-TNF-a活性蛋白
TNF-a是内源性的热原,能够引起发热、凋亡细胞死亡、炎症和抑制肿瘤发生。据报道,TNF-a可以抑制A549细胞的增殖和诱导凋亡,在刺激后细胞上清液中IL-1的浓度会增加。
实验方法:因此,A549细胞在DMEM孵化TNFa(1 ng / mL,10 ng / mL)2 h,4 h,8 h,24小时、48小时,然后细胞被倒置显微镜观察,IL-1β被ELISA检测细胞上清液中。
实验结果:用TNF-a (10ng/mL)孵育72h后观察细胞凋亡(见图一)。
A B
Figure 1. Effect of TNF-a on A549 cells.
(A) A549 cells cultured in DMEM, stimulated with 10ng/mL TNF-a for 72h;
(B) (B)Unstimulated A549 cells cultured in DMEM for 72h.
与TNF-a孵化后(10 ng / mL)8 h,IL-1β显著增加的细胞上清液(见表一)。
Sample (cell supernatant of A549 cells) | O.D. value | Corrected | Concentration of IL-1β (ng/mL) |
stimulated with TNF-β (10ng/mL) | 1.210 | 1.046 | 52.0 |
unstimulated | 0.187 | 0.023 | 4.9 |
Table1.Effect of TNF-β on A549 cells by ELISA.
STD (ng/mL) | O.D. value | Corrected |
150 | 3.363 | 3.199 |
75 | 1.630 | 1.466 |
37.5 | 0.974 | 0.810 |
18.75 | 0.442 | 0.278 |
9.375 | 0.271 | 0.107 |
4.6875 | 0.180 | 0.016 |
2.34375 | 0.168 | 0.004 |
The standard curve was demonstrated as follows: