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Western blot analysis of Rabbit IgG heavy chain(Fc fragment) on Rabbit IgG (positive) with Mouse anti-Rabbit IgG heavy chain(Fc fragment) antibody (HA601651) at 1/50,000 dilution.
Lysates/proteins at 10 µg/Lane.
Exposure time: 1 minute; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA601651, 1/50,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Observed band size: 50 kDa
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☑ Relative expression (RE)
Western blot analysis of Rabbit IgG heavy chain(Fc fragment) on Mouse IgG (negative) with Mouse anti-Rabbit IgG heavy chain(Fc fragment) antibody (HA601651) at 1/50,000 dilution.
Lysates/proteins at 10 µg/Lane.
Exposure time: 1 minute; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA601651, 1/50,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
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☑ Relative expression (RE)
Western blot analysis of Rabbit IgG heavy chain(Fc fragment) on Human IgG (negative) with Mouse anti-Rabbit IgG heavy chain(Fc fragment) antibody (HA601651) at 1/50,000 dilution.
Lysates/proteins at 10 µg/Lane.
Exposure time: 1 minute; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA601651, 1/50,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
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Indirect ELISA analysis of Rabbit IgG heavy chain(Fc fragment) was performed by coating wells of a 96-well plate with 50 µL per well of Fc fragment of Rabbit lgG / F(ab')₂ fragment of Rabbit IgG / Human IgG diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 1%BSA blocking buffer for 1 hour at 37℃, and incubated with 50 µL per well of Rabbit IgG heavy chain(Fc fragment) recombinant Mouse monoclonal antibody serial diluted starting from a concentration of 4µg/mL for 45 minutes at 37℃. The plate was washed and incubated with 50 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1/80,000 for 30 minutes at 37℃. Detection was performed using an Ultra TMB Substrate for 10 minutes at 37℃ in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
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