Human IgE Antibody (F4-#16)

Western Blot: Human IgE Antibody (F4-#16) [NBP1-78578]

Western Blot: Human IgE Antibody (F4-16) [NBP1-78578] - Analysis of 0.5ug of purified human IgE, IgM, IgA and IgG per well and 10ul of PageRuler Prestained Protein Ladder per well.

ELISA: Human IgE Antibody (F4-#16) [NBP1-78578]

ELISA: Human IgE Antibody (F4-16) [NBP1-78578] - Direct ELISA analysis of human IgE was performed by coating wells of a 96-well plate with100ul per well of human IgE, IgM, IgA, or IgG, diluted in carbonate/bicarbonate buffer at concentrations ranging from 5-5000ng/ml, and incubating overnight at 4C. The plate was washed and blocked with StartingBlock T20 (TBS) Blocking Buffer for at least 30 minutes at room temperature, and then incubated with 100ul per well of a human IgE monoclonal antibody at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, and incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:8000 for at least 30 minutes at room temperature. Detection was performed by adding 100ul per well of 1-Step Ultra TMB substrate and incubating for a few minutes at room temperature. The reactions were then stopped with TMB Stop Solution. Absorbances were read on a spectrophotometer at 450-550nm.

ELISA: Human IgE Antibody (F4-#16) [NBP1-78578]

ELISA: Human IgE Antibody (F4-#16) [NBP1-78578] - 96-well plate coated with 100ul/well of goat anti-human IgG (H+L) antibody (1ug/ml, carbonate/bicarbonate buffer), incubated overnight at 4C. Plate washed & blocked with StartingBlock T20 (TBS) for 30 mins at RT, then 100ul /well of human IgE (1X PBS, 1-1000ng/ml, left), & normal human serum or serum from asthma patient on prescription medications (control serum, right, 1X PBS, 1:5 - 1:5000), added to plate & incubated for 1 hr at RT. Plate washed with 150ul /well of 1X TBST, & 100ul (1ug/ml) human IgE monoclonal antibody added to wells, & incubated for 1 hr at RT. Plate washed & incubated with 100ul /well of an HRP-goat anti-mouse IgG Fc secondary antibody at 1:8000 for 30 mins at RT. Detection performed by adding 100ul /well of 1-Step Ultra TMB substrate & incubating for a few min at RT. Reactions stopped with TMB Stop Solution. Absorbances were read on a spectrophotometer at 450-550nm.