ELISA
Microtiter plate wells were coated with 100 ng/well coating antigen and incubated with serial dilutions of the antibody. Following HRP-conjugated secondary antibody and substrate development, OD630 was measured. The absorbance decreased progressively with increasing antibody dilution, yielding a sigmoidal titration curve that demonstrates the high antigen-binding activity of the polyclonal antibody.
DB
Dot blot analysis was performed using Anti-Phospho-Insulin Receptor(Tyr1355) antibody to detect two peptide samples: lane 1 contains the phospho-peptide , and lane 2 contains the non-phospho-peptide.
Transfer the proteins onto a NC membrane (
OM790001), and block it with TBST (
OM750016) containing 5% BSA(OM297470) for 1 hour. Dilute the primary antibody with the antibody diluent (
OM750012) at a ratio of 1:1000, and incubate it at room temperature for 2 hours. Wash the membrane 3 times with TBST (
OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (
OM643487), at a ratio of 1:10000 and incubate for 1 hour. Wash the membrane 3 times with TBST (
OM750016) again, 5 minutes each time. Use ECL (
OM625701) for luminescence. Exposure time: 30 s.