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Supplemental Product Information:

Product Information

Volume/Quantity : 0.5 mg x 3 vial
Conjugation : Peroxidase from Horseradish (HRP) was conjugated to Sulfo-SMCC (Product No. S0883).
Product Form : Horseradish Peroxidase Maleimide Conjugate. Lyophilized in 10 mM sodium phosphate, 0.115 mM NaCl, 40 mM sucrose, pH 6.6.
Preservative : None

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.

Protocol

1. Firstly return the vial to room temperature.
2. 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.
3. 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.
4. 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.
5. 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.

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