Montage Antibody Purification Kits

Designed for fast and easy antibody purification from serum, ascites or cell culture supernatants.

Overview

Specifications

Ordering Information

Montage Antibody Purification Kits Clear Sorting & Filtering
Catalogue Numbericon Descriptionicon Pack Sizeicon
LSK2ABA20Montage Antibody Purification Kit with PROSEP-A media 2 columns (20 purifications) Show Pricing & Availability
LSK2ABA60Montage Spin Columns with PROSEP-A media 6 columns (60 purifications) Show Pricing & Availability
LSK2ABG20Montage Antibody Purification Kit with PROSEP-G media 2 columns (20 purifications) Show Pricing & Availability
LSK2ABG60Montage Spin Columns with PROSEP-G media 6 columns (60 purifications) Show Pricing & Availability
SCGP00525Steriflip-GP Sterile Centrifuge Tube Top Filter Unit 25 Show Pricing & Availability
UFC903008Amicon Ultra-15 Centrifugal Filter Unit 8 Show Pricing & Availability
UFC903024Amicon Ultra-15 Centrifugal Filter Unit 24 Show Pricing & Availability
UFC903096Amicon Ultra-15 Centrifugal Filter Unit 96 Show Pricing & Availability

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    Documentation

    Data Sheet

    Title
    Montage Antibody Purification Kits

    Posters

    Title
    Poster: Rapid, Ultrafiltration-based Method for Purification of Monoclonal Antibodies from Hybridoma Supernatants

    References

    Reference overview
    Antibody Purification with Affinity Spin Columns
    Kavonian, Mark,BioScience Technology, July 2003
    BioScience Technology, July 2003  2003

    Considerations During Development of a Protein A-Based Antibody Purification Process
    Iyer et al.,Biopharm, January 2002, pp 14-20
    Biopharm, January 2002, pp 14-20  2002

    A sensitive enzyme linked immunoadsorbent assay (ELISA) for the detection of staphylococcal prtein A (SpA) present as a trace contaminant of murine immunoglobulins purified on immobilized protein A
    Miguel A.J. Godfrey, Piotr Kwasowski, Roland Clift and Vincent Marks
    Journal of Immunological Methods, 149(1992) 21-27, 1992  1992

    FAQ

    QuestionAnswer
    Can I use a fixed angle rotor or a swinging bucket rotor with the Prosep A and Prosep G spin columns?Swing bucket rotors are recommended because they provide uniform flow through the resin. For optimal performance with a fixed angle rotor, ensure that the orientation of the Prosep A spin coulmn is the same for all steps. Also please note that the maximum sample loading volume for a fixed angle rotor is ten ml versus twenty ml with a swinging bucket rotor.
    How do I monitor the purity of isolated antibodies?Purity is best measured by gel electrophoresis. When analyzed by SDS-PAGE under non-reducing conditions, IgG anitbodies should give a single protein band at about 160-170 kDa. On reduction with DTT or 2-mercaptoethanol, two or more bands will be seen corresponding to the individual heavy chains (50-55 kDa) or light chains (25 kDa).
    Are there any critical starting conditions for use of the Prosep A spin columns?For optimal performance, the pH should be above 8.0.
    How can I regenerate the Prosep G spin columns?We recommend that you wash the plugs with 10 ml of Elution Buffer B2 (pH2.5) by centrifuging the spin columns at 500 xg for 5 minutes. Re-equilibrate the plugs with 5 ml of Binding Buffer A by centrifuging the spin columns at 500 xg for 2 minutes. For immediate re-use wash with 5 ml of Binding Buffer A or for later use store the plugs without their end caps in 5 ml of Binding Buffer A in a 15 ml screw-capped tube.
    What are the critical starting conditions for the Prosep G spin columns?For optimal performance the pH should be above 8.0.
    Do I need to control the salt concentration when using the Prosep G spin columns?Use 0.1-0.5 M salt to reduce non-specific adsorption.
    The sample does not flow easily through the Prosep A and Prosep G spin column. what could be the cause?There could be 3 causes:

    1. The media is clogges with particulates, most likely because the sample was not pre-filtered, or because the serum is particularly thick.
    2. If the spin columns are not stored at 2-8 C microbial growth could occur in the media plug causing a restriction of flow.
    3. The g force for the sample loading step should be increased up to 150 x g.
    The target protein doesn't seem to be eluting from the Prosep G spin column. Could there be a cause?The pH of the elution buffer may be inccorect. If it is not acidic enough, the target protein will not desorb from the spin column. It would be advisible to prepare a new solution and check the pH.
    The recovery of my target protein from the Prosep Aand Prosep G spin columns is low, what could be the cause?The binding of antibodies to the Prosep A and Prosep G spin columns is attributed in part to hydrophobic forces. Chaotropic salts can be added to the elution solution reducing the strength of the hydrophobic interactions.
    Can the antibody purification with the Montage Antibody Purification Prosep A/G columns be done with gravimetric flow instead of centrifugation?No. There is not enough force to run via gravimetric flow. These devices designed for centrifugation.

    User Guides

    Title
    Montage Antibody Purification Kit and Spin Columns with PROSEP-A Media
    Montage Antibody Purification and Spin Columns with PROSEP-G Media

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    Categories

    Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Agarose Beads for IP & Antibody Purification
    Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Agarose Beads for IP & Antibody Purification
    Life Science Research > Protein Sample Preparation > Protein Purification > Agarose Bead Affinity Purification > Agarose Beads for IP & Antibody Purification