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Micro Peptide Coupling Kit


Polyacrylamide resin for coupling sulfhydryl-containing ligands.

Micro Peptide Coupling Kit

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.

Highlights:

  • 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

Product Details:

Protocol summary for the Thermo Scientific Micro Peptide Coupling Kit
Protocol summary for the Thermo Scientific Micro Peptide Coupling Kit. Learn more about sulfhydryl-reactive immobilization supports and about haloacetyl crosslinking chemistry.

Gel analysis of anti-insulin purified using the Thermo Scientific Micro Peptide Coupling Kit
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 (Part No. 26681).

 
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).

Anti-insulin purified using the Thermo Scientific Micro Peptide Coupling Kit maintains activity
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.

Related Resources:

Review of affinity purification
Review of covalent immobilization methods
Sulfhydryl-reactive crosslinker chemistry

Related Products:

UltraLink Iodoacetyl Resin – stand-alone resin used in the Micro Peptide Kit
SulfoLink Immobilization Kit – larger scale kit for peptide immobilization to beaded agarose resin
Micro Protein Coupling Kit – for protein coupling through primary amines (lysine residues)


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
20485 UltraLink Iodoacetyl Micro Peptide Coupling Kit, 0.1mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 25-250µg of peptide or protein per microcentrifuge column; Includes buffers for 20 affinity purifications

Kit Contents:
UltraLink Iodoacetyl Spin Columns, 0.1mL, 10 columns
Coupling Buffer, 100mL
L-Cysteine·HCl, 100mg
Wash Solution, 25mL
PBS Pack (makes 500mL), 1 pack
IgG Elution Buffer, 50mL
Collection Tubes, 200 tubes
10-column kit Product Instructions for product #20485 UltraLink Iodoacetyl Micro Peptide Coupling Kit, 0.1mL MSDS for product #20485 UltraLink Iodoacetyl Micro Peptide Coupling Kit, 0.1mL Certificate of Analysis for product #20485 UltraLink Iodoacetyl Micro Peptide Coupling Kit, 0.1mL $385.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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