Anti flag ae005

Antisera specific to viral GP1, GP2, and NP were generated by immunizing BALB/c mice with purified viral proteins. The anti-STT3A (HPA030735) and anti-STT3B (HPA036646) antibodies were purchased from Sigma-Aldrich (USA). The anti-Strep (catalog no. A00626) antibody was purchased from GenScript (China). The anti-V5 (AB3792) antibody was purchased from Merck Millipore (Germany). The anti-MAGT1 (17430-1-AP) and anti-6×His (66005-1-Ig) antibodies were purchased from Proteintech (China). The anti-TUSC3 (A12571), anti-GAPDH (AC002), and anti-Flag (AE005) antibodies were purchased from ABclonal (China). MagStrep XT beads (2-4090-002) and buffer BXT (2-1041-250) were purchased from IBA Life Sciences (Germany). NGI-1 (HY-117383) was purchased from MedChemExpress (China).

RCC cells were collected and lysed with RIPA (C1053, Beijing Pulley Gene Technology Co., Ltd., Beijing, China) containing protease inhibitor cocktail (HY-K0010, MCE, Shanghai, China). The obtained proteins were incubated with primary antibodies (anti-Flag, AE005, 1:100, mouse antibody; anti-Myc, AE070, 1:50, rabbit antibody, Abclonal, Woburn, MA, USA) at 4°C overnight. Protein A/G-agarose beads (36403ES08, Yeasen, Shanghai, China) were added for 4 h to obtain immunoprecipitates, which were then purified at least three times in a lysis buffer. Purified complexes were treated by 2 × SDS loading buffer, boiled, electrophoresed and assayed by Western blot.