Analyzing transcription factor activity using near infrared fluorescent bridge polymerase chain reaction

摘要: This study has developed a new method, near infrared fluorescent bridge polymerase chain reaction (NIRF-bPCR), for analyzing transcription factor (TF) activity. method was first used to detect the activity of purified nuclear kappa B (NF-κB) p50. The results demonstrated that this could quantitatively p50 protein at less than 115ng (∼ 2320fmol), and detection limit reached as little 6.94ng 140fmol) protein. then TF in cell extracts. revealed specifically NF-κB HeLa Finally, activities multiple TFs sample. showed six TFs-NF-κB, AP-1, TFIID, CREB, NF-E2, p53-in TNFα-induced -uninduced Calculation fold induction NF-κB, AP1 were activated by TNFα cells, agreement with other methods. Therefore, provides tool also NIRF-bPCR may be detecting DNA molecules.