Goat anti mouse lgg hrp

Proteins were extracted using RIPA lysis buffer (strong) (Beyotime, China) from 2-week-old seedlings grown hydroponically on medium containing different N concentrations of 0.02 mM, 0.2 mM, and 2 mM KNO₃ without salt as a control or with 100 mM NaCl. For western blot analysis, proteins were electroblotted from a 10% acrylamide gel to a nitrocellulose membrane (Immobilon-P, Millipore, Bedford, MA, USA) after SDS‒PAGE separation. The antibodies used in western blotting were as follows: anti-GFP antibody (M20004, Mouse mAb, Abmart, Shanghai, China), 1:1000; anti-ACTIN antibody (M20009, Mouse mAb, Abmart, Shanghai, China), 1:1000; and goat anti-mouse lgG-HRP (M21001, Abmart, Shanghai, China), 1:5000. An Image Quant LAS 4000 (GE, USA) CCD camera system was used to quantify band intensity with the Super Signal West Femto Trial Kit (Thermo, Rockford, IL, USA).