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.

 

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