Rabbit anti goat igg cy3 antibody

For immunofluorescence, MAC-T cells were cultured and treated in a 6-well plate as recently described (Ma et al., 2020) (link). The cells were incubated with the primary antibody against p-p65, followed by incubation with rabbit anti-goat IgG/Cy3 antibody (Bioss, Beijing). The nuclei were stained blue with 4′, 6-diamidino-2-phenylindole and observed with a fluorescent inverted microscope (Leica).
For the ELISA test of p65 nuclear translocation, MAC-T cells were cultured and treated as previously described in a 6-well plate. The homogenate of mammary gland samples was centrifuged at 4°C and 3,000 × g for 10 min. Nuclear protein of the cultured cells and tissues was extracted using nuclear protein extraction kits and quantified using bicinchoninic acid protein quantification kits (Vazyme). The contents of p-p65 in nuclear were assessed using commercial ELISA kits (Youxuan). Each ELISA reaction was performed in triplicate. The optical density values were measured at 450 nm in xMarkTM (BIO-RAD).