Technical Reference #1826

1826.

Hemokinin-1 Activates the MAPK Pathway and Enhances B Cell Proliferation and Antibody Production Wei Wang; Qian Li; Jun Zhang; Hounan Wu; Yanhui Yin; Qing Ge; and Yu Zhang, Peking University Health Science Center, Journal of Immunology, 184(1826), (2010)
Link To Paper

Materials & Methods:
Calcium flux- B cells were added into poly-L-lysine–coated glass bottom microwell dishes designed for confocal microscopic examination (MatTek Ashland MA) at a density of 1 3 106 cells/ml and allowed to attach for 12 h. The cells were then loaded with Fluo-3 acetoxymethyl ester (Biotium Hayward CA) at 37°C for 60 min. Intracellular Ca2+ concentration was measured by monitoring the fluorescence of individual cells at 525 nm using a laser scanning confocal microscope (DM IRB Leica Microsystems Deerfield IL). Baseline fluorescence were assessed for 30 s before adding anti-IgM (200 ng/ml) HK-1 (0.5 mM) or anti-IgM plus HK-1. Image recording continued for additional 10 min after stimulation. Ca2+ concentrations were expressed as the average fluorescence intensity of 15–20 cells randomly picked up from at least two fields.

Microscopic Technique
Laser Scanning Confocal Microscope

Cell Type(s)
Splenic B cells