β-Lactoglobulin Influences Human Immunity and Promotes Cell Proliferation

Figure 4

Purification of the LG receptor. (a) SDS-PAGE: lane I: protein ladder, lane II: LG receptor, and lane III: LG receptor with a reducing agent (β-ME); (b) Western blot. The receptor was purified using a LG affinity column. Samples from the column were analyzed by SDS-PAGE and Western blot analysis. A protein (later confirmed as the LG receptor) with a molecular weight of approximately 150 kDa was detected using a specific antibody. (c) Hybridoma cells were incubated with (1) IgG (1 mg) purified from normal rabbit serum (control) or (2) rabbit polyclonal antibodies specific to the LG receptor (1 mg). After labeling these two antibodies with FITC, the FITC-conjugated antibodies were incubated with the cells at 4°C for 30 min. We analyzed the localization of the FITC-labeled proteins using confocal microscopy (magnification = 1000x). Only rabbit polyclonal antibodies specific to the LG receptor (2) were detected on the cell membrane (green colored fluorescence). These data further confirmed that LG receptors were located on the surface of the membrane.

(a)

(b)

(c)