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Hemocyanin, Maleimide Activated, Low Endotoxin, Highly Soluble, Keyhole Limpet, Megathura crenulata,
[H-1186]
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Full Name: Hemocyanin, Maleimide Activated, Low Endotoxin, Highly Soluble, Keyhole Limpet, Megathura crenulata; [KLH, Maleimide Activated]
Description: High solubility form of Keyhole Limpet Hemocyanin (KLH) activated with succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-caboxylate and formulated to use directly with cysteine containing peptides or haptens that contain a free SH group.
KLH is a copper-containing protein isolated from the mollusc Megathura crenulata and a potent immunoactivator commonly used as a carrier in the conjugation of peptides for antibody production. The preparation of KLH used to produce the Maleimide Activated form is chromatographically purified and treated to reduce endotoxins for immunization protocols.
Purity: Activation: 150-300 maleimide groups per molecule of KLH (mol. wt. ~ 8 x 10 to the 6. Each lot of Maleimide Activated KLH is assayed for total number of maleimide groups by titration with 5-((2-(and-3)-S-(acetylmercapto)succinoyl)amino)fluorescein (SAMSA fluorescein).
Physical Purity: 90% by size exclusion chromatography as a mixture of high and low molecular weight forms; Endotoxin: ≤12 EU/mg; Copper to protein ratio: ≥1.5 x 10-3.
Reconstition: Reconstitute with 1 mL of 10 mM phosphate buffer at 4ºC containing 1 mM EDTA, pH 7.0. Aliquots may be frozen immediately after reconstitution and are stable for approximately 3 months.
Reaction directions: Add SH-peptide dissolved in phosphate buffer + 1 mM EDTA to the reconstituted KLH-Activated solution using a 10X molar excess of SH-peptide. Assuming a maximum of 300 reaction sites per molecule of KLH and an average mol. wt. of 8 x 106 for KLH, the following calculation can be used to determine the amount of SH-peptide to add per mg of KLH. Peptide required = (0.13 nMole KLH /mg protein) x (300 sites/molecule of KLH) x (5-10 fold excess peptide) or 180-360 nMoles (peptide) per mg KLH.
Conjugation at room temperature is nearly complete after one hour. The reaction may be allowed to proceed overnight at 4ºC. Any remaining sites may be quenched with 5 mM cysteine for 30 minutes followed by gel filtration to remove unconjugated reaction products.
Example:
• Example: For a peptide having a molecular weight of 1000, the milligrams of peptide required is 0.18-0.36 mg per mg KLH
Formulation: Lyophilized from 5% sucrose (stabilizer), 10 mM sodium phosphate, pH 7.0 at a protein concentration of approximately 10 mg per mL this formulation is composed of both high molecular weight aggregates and low molecular weight monomeric units for direct use in peptide conjugation.
Storage: -20C.
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$225.00
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Note: All products sold by A.G. Scientific are for Laboratory or Manufacturing purposes only. They are not intended for Human Drug, Food Additive, Clinical or Household use.
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