Polyacrylamide resin for coupling sulfhydryl-containing ligands.
The Thermo Scientific Micro Peptide Coupling Kit enables covalent immobilization of cysteine-containing peptides for use in antibody purification and other affinity protocols.
The Micro Peptide Kit is for immobilizing small amounts (25 to 250µg) of sulfhydryl-containing peptides (e.g., cysteine-terminated peptides) onto a beaded porous resin to create a small, reusable, microcentrifuge affinity column. The coupling and affinity purification procedures are optimized for small sample volumes (200 to 300µL). Wash and elution steps are achieved rapidly and efficiently with the convenient microcentrifuge spin columns. Each kit contains sufficient reagents for 10 coupling reactions and 20 affinity purifications. The kit is ideal for immobilizing peptide antigens that contain a terminal cysteine residue for use in purifying specific antibodies from small serum, ascites or culture supernatant samples.
Optimized for small scale – ideal for coupling small amounts (25 to 250µg) of sulfhydryl-containing peptide or protein for purification of specific antibodies from crude serum
High-performance affinity resin – uses durable, polyacrylamide-based UltraLink Iodoacetyl Resin for specific reaction to sulfhydryl groups
Efficient coupling chemistry – immobilization efficiency greater than 85% (as measured with 1 hour reaction using insulin, calcitonin and osteocalcin peptides)
Fast and easy to use – perform wash and elution steps using a microcentrifuge
Reusable – use prepared peptide resin several times with no significant loss of capacity
Efficient purification of anti-peptide antibody from crude serum using the Thermo Scientific Micro Peptide Coupling Kit. Insulin peptide (245µg) coupled in a kit spin column was used to purify anti-insulin from 40µg of bovine insulin anti-serum. Eluted protein was separated by SDS-PAGE (4-20% gel) and stained with Thermo Scientific GelCode Blue Stain Reagent (Product No. 24590).
Lane 1: 40µg bovine insulin anti-serum (crude serum).
Lane 2-7: Alternating non-reduced and reduced fractions (A = whole IgG; B = heavy chain; C = light chain).
Lanes 2 & 3: Eluted fraction 1 (10µL each).
Lanes 4 & 5: Eluted fraction 2 (10µL each).
Lanes 6 & 7: Eluted fraction 3 (10µL each).
Lane 8: MW Marker (Product # 26681).
Anti-insulin purified using the Thermo Scientific Micro Peptide Coupling Kit maintains activity. Increasing concentrations of purified bovine insulin was transferred to nitrocellulose membrane, probed with kit-purified anti-insulin and detected by chemiluminescence with Thermo Scientific SuperSignal West Pico Substrate (Part No. 34080) after incubation with the appropriate secondary antibody HRP conjugate.