Description: HRP Maleimide Conjugate is designed to facilitate the conjugation of the HRP to proteins, peptides and ligands that contain a free sulfhydryl (-SH) groups. Activated HRP contains 1–3 maleimide groups for each HRP molecule.
Product Information: Volume/Quantity : 0.5 mg x 3 vial Conjugation : Peroxidase from Horseradish (HRP) was conjugated to Sulfo-SMCC (Product M123456). Product Form : Horseradish Peroxidase Maleimide Conjugate. Lyophilized in 10 mM sodium phosphate, 0.115 mM NaCl, 40 mM sucrose, pH 6.6. Preservative : None
Protocol: Firstly return the vial to room temperature. Reconstitute the contents of the vial with 100μL of buffer or water to obtain a 5 mg/mL solution of HRP Maleimide. Note: Do not vortex or heat the HRP Maleimide. Dissolve the sulfhydryl compound to be conjugated at 2-5 mg/ml in the buffer. Note: We recommend that the thiol-containing molecule is dissolved in the pH range 6.5-7.5 buffer. 0.1M sodium phosphate, 0.15M NaCl, 10mM EDTA at pH 7.2 is commonly employed. Maleimides react with sulfhydryls at pH 6.5-7.5 to form stable thioether bonds. EDTA may help to prevent metal catalysed oxidation of thiols. Use of a 4:1 (HRP Maleimide:antibody) molar ratio in the conjugation reaction usually results in high-activity conjugates. This vial contains 0.5 mg of HRP Maleimide Conjugate and can effectively modify 0.5 mg of immunoglobulin. Immediately mix the thiol-containing molecule and HRP Maleimide solutions and allow them to react for 2 hours at room temperature. To increase HRP incorporation, extend reaction time up to 12 hours. If desired, desalt, dialyze or purify the conjugate.
Storage: Store at −20 °C to −80 °C. Reconstitute this reagent immediately before use. Do not prepare stock solutions for storage. Discard any unused reconstituted reagent. |