Carrier Proteins (KLH, BSA etc) and Fusion protein (GST) Agarose
 for Affinity Purification of Antibodies

Antibodies to small peptide or haptens are generally raised by coupling to large carrier proteins such as Keyhole limpet Hemocyanin (KLH), Bovine serum albumin (BSA), Ovalbumin and thyroglobulin etc.  However, antibodies are produced to the carrier proteins (KLH, BSA etc) and the small peptide/hapten.  Many recombinant proteins are expressed and purified from E. coli.  Often, it is difficult to remove the contaminating E. coli proteins.  Antibodies made to proteins containing the E. coli protein contaminants can be removed by immunoadsorption over the E. coli protein-agarose column.  Similarly, antibodies made to GST-fusion proteins can be immunoadsorbed over the GST-Agarose to remove anti-GST antibodies.  
Anti-carrier proteins/fusion proteins may give non-specific signals in various immunoassays.  There are 2 common ways to clean up the crude antiserum.  

1. Selectively purify antibodies to the antigen by making an antigen-agarose affinity column and subsequent antibody purification.
2. Remove the carrier protein antibodies using commercially available Carrier-protein Agarose (KLH/BSA-Agarose) using solid phase immunoaffinity column chromatography.
3. Couple specific antigens or proteins to purify or remove antibodies using Pre-activated Agarose gels.

Carrier/Fusion Protein Agarose-Ordering Information

Keyhole Limpet Hemocyanin, KLH-Agarose, Cat # KLH11-G (for the removal of KLH antibodies)

E.Coli Proteins-AgaroseCat # EC11-G (for the removal of E. coli protein’s antibodies)

GST protein-Agarose, Cat # GST15R-AS 
(for the removal of GST antibodies or to purify GST binding proteins

Chicken Egg-Ovalbumin-Agarose, Cat # OVA15-AS 
(for the removal/or purification of ovalbumin antibodies)

Bovine Serum Albumin (BSA)-Agarose, Cat # BSA15-AS 
(for the removal/or purification of ovalbumin antibodies)

Pre-Activated Agarose gels-Ordering Information
A variety of easy to use kits are available to coupled peptides and proteins to agarose.  No complicated reagents or chemicals are required.  All reagents are provided and the coupling time is usually 2-4 hrs at room temp. 
Click here for catalog & ordering
ItemName                   CatalogNumber       Product Description, Quantity/SIZE  
C-Link Agarose               110100-CG                Preactivated C-Link-Affinity Gel Sepharose (for Cys-containing peptides/proteins),  5 ml 
N-Link Agarose               110200-NG                Preactivated N-Link-Affinity Gel Agarose (for NH2-containing peptides/proteins)       5 ml 
Affinity purification kit      #110300-AF              A kit containing all affinity purification buffers for 10-20 antibody purifications  

Custom conjugation of peptides/protein to Agarose
Cat # AFFI-2, Custom conjugation of peptide to agarose, $295 per peptide (couple ~2-5 mg peptide) 
Affinity matrix is supplied in a ready-to-use mini column.  Time ~2-4 working days.   We also offer complete affinity purification package Cat # AFFI-I for the preparation of affinity matrix, purification of antibodies, and ELISA testing.  Time 3-5 working days.   This package typically includes: Coupling of peptide/proteins (up to 2-5 mg peptide to Agarose), purification of any 1 bleed or ~ 15 ml antiserum; ELISA testing of both unpurified and affinity pure antibodies, and supply of affinity column. We also monitor the antibody activity in the unbound fraction to make sure antibodies are not overloaded and wasted.