[Detection of nuclear factor kappa B by immunochemical staining and image analysis].
OBJECTIVE
To find a relatively simple and safe method for quantitative assay of nuclear factor (NF)-kappa B activity.


METHODS
Immunochemical staining of NF-kappa B/p65 subunit with its antibody was performed in human umbilical vein endothelial cells stimulated with advanced glycation end products-modified human serum albumin (AGE-HSA). The ratio of p65 subunit staining in the nuclei and cytoplasm was determined by means of imaging analysis, and the results were compared with those of electrophoretic mobility shift assay (EMSA).


RESULTS
NF-kappa B could be activated by AGE-HSA in a time- and dose-dependent manner, and the results obtained from immunochemical staining were consistent with those from EMSA.


CONCLUSIONS
p65 immunochemical staining and subsequent image analysis is feasible in the quantitative detection of NF-kappa B activation.
Zhang
Gui-lin
GL
Department of Nephrology, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Liu
Shang-xi
SX
Deng
Yan
Y
Shen
Hong
H
D024202:Electrophoretic Mobility Shift Assay; D017127:Glycation End Products, Advanced / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D016328:NF-kappa B / Q000032:analysis*
D017127:Glycation End Products, Advanced; D016328:NF-kappa B