Improve the performance of antibody conjugates by removing excess free enzyme (HRP or AP).

Thermo Scientific Pierce Conjugate Purification Kit is a gentle affinity chromatography system for purifying horseradish peroxidase (HRP) or alkaline phosphatase (AP) antibody conjugates from the non-conjugated enzymes following enzyme-antibody conjugation reactions. The metal chelate agarose column binds to IgG and IgG-enzyme conjugates but allows non-conjugated enzyme (HRP or AP) to to pass through. A gentle, non-denaturing and elution buffer recovers the fully active IgG-HRP or IgG-AP conjugate, ready for use in immunodetection methods.

Purification of IgG-HRP removes excess non-conjugated enzyme and recovers fully active HRP conjugated antibody. The solution resulting from a somewhat inefficient conjugation reaction (5 mg total protein) was loaded to the nickel-charged column and purified using the Conjugate Purification Kit. Test samples from various time-points in the procedure were tested for HRP activity by in a microplate assay using ABTS (Product # 37615) as a substrate.
 

Advantages of making and using purified antibody conjugates:

• Reduced background for blotting and ELISA
• Fewer washes needed to remove background
• Increased sensitivity of assay system with more active reagent
• Improved signal-to-noise ratio of conjugate

  Product #	Description	Pkg. Size
  44920	Pierce Conjugate Purification Kit Sufficient reagents for 10 antibody conjugate purifications. Kit contains: Immobilized Iminodiacetic acid, 1 x 2 ml column Polyacrylamide Desalting Column, 1 x 10 ml column Pierce Column Activator Buffer, 25 ml Pierce Elution Buffer, 35 ml BupH Tris Buffered Saline Packs, 2 each BupH Phosphate Buffered Saline Pack, 1 each Activity Preservation Solution, 1 ml	Kit