Human IgM Antibody (P5E2)

Western Blot: Human IgM Antibody (P5E2) [NBP1-78603]

Western Blot: Human IgM Antibody (P5E2) [NBP1-78603] - Analysis of 1ug of purified Human IgM, Human IgA, and Human IgG and 10ul of PageRuler Prestained Protein Ladder per well.

Flow Cytometry: Human IgM Antibody (P5E2) [NBP1-78603]

Flow Cytometry: Human IgM Antibody (P5E2) [NBP1-78603] - Analysis of surface IgM expression was performed on Ramos (surface and cytoplasmic IgG/lambda-expressing B cells), Raji (cytoplasmic IgG/kappa-expressing B cells), and Jurkat (T cells) cell lines. Cells were stained with a mouse anti-human IgM monoclonal antibody or mouse IgG1 isotype control (black histogram), each at a concentration of 1ug/ml. After incubation of the primary antibody on ice for 1 hour, the cells were stained with a FITC-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:500 for 30 minutes on ice. The representative 10,000 cells were obtained for each sample.

ELISA: Human IgM Antibody (P5E2) [NBP1-78603]

ELISA: Human IgM Antibody (P5E2) [NBP1-78603] - Direct ELISA analysis of various immunoglobulin (Ig) isotypes was performed by coating wells of a 96-well plate with 100ul per well of Human IgM, Human IgG, or Human IgA diluted in carbonate/bicarbonate buffer, starting at a concentration of 50ng/ml and serially diluting 2-fold to a concentration of 0.05ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse anti-human IgM monoclonal antibody at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.