Overview

  • Product nameAntibody Concentration Kit
  • Product overview

    Antibodies are sometimes only available at low concentrations and often contain low molecular weight substances that interfere in labeling reactions with enzymes, biotin, streptavidin and fluorophores. The Antibody Concentration Kit allows for the quick and easy concentration of antibodies and proteins. The kit can also be used to reduce the concentration of many unwanted additives often found in antibody formulations such as azide, glycine or tris.

    The Antibody Concentration Kit method utilizes a simple spin column to easily and quickly remove excess buffer from the antibody thereby providing a more concentrated antibody solution. The Antibody Concentration Kit also allows the experimenter to perform a simple buffer exchange to transfer the antibody.

    The components of Ab102778 are fully compatible with our Antibody conjugation kits however they are not compatible with our GOLD Antibody conjugation kits. To concentrate antibodies for use with our GOLD conjugation kits, please use our Gold antibody concentration kit (ab204911).

     

  • Notes

    Concentration of Antibody Solution

    1. Add antibody to spin cartridge.

     

    • Spin for 1 to 3 minutes* in a microfuge at max speed of 15000g to reduce the buffer volume in the spin cartridge to between 50 and 100ul. It is advisable not to spin the antibody dry as reconstitution of the antibody will be difficult and significant antibody loss and degradation may occur.

     

     

    • Repeat steps 1 to 2 as many times as is necessary to process the entire antibody to the desired concentration. It may be necessary to discard the excess buffer collected in the collection tube between spins.

     

     

    • Recover the concentrated antibody from the spin cartridge. NB. It is advisable not spin the antibody dry as reconstitution of the antibody will be difficult and significant antibody loss and degradation may occur.

     

    Buffer Exchange Using Spin Column Assembly

    1. Add up to 0.5ml antibody to spin cartridge.

     

    • Spin for 1 to 3 minutes* in a microfuge at maximum speed to reduce the buffer volume to 100ul.

     

     

    • Discard the excess liquid in collection tube.

     

     

    • Add 400ul conjugation buffer to the antibody in the spin cartridge.

     

     

    • Spin for 1 to 3 minutes* in a microfuge at maximum speed to reduce buffer volume to 100ul.

     

     

    • Discard the excess liquid in collection tube.

     

     

    • Repeat steps 1 to 6 at least 5 times to exchange antibody buffer.

     

     

    • Recover antibody from the spin cartridge.

     

    *Spin times will vary depending on buffer composition, starting volume and centrifuge speed.

     

    Note Each cycle leads to a reduction in the concentration of low molecular substances. By performing as many as 5 repeat steps the concentration of small molecules such as glycine and Tris will be reduced 2500 fold. However, the concentration of proteins such as BSA will be unchanged.

     

    To remove unwanted proteins see ab102784.

     

    Storage of Antibody

    Store at 4°C. Other storage conditions (e.g. frozen at -70°C may also be satisfactory). The sensitivity of any particular antibody to freeze thaw should be determined by experimentation on small aliquots.

     

    Test for Protein

    Wherever possible protein values should be determined using an absorbance at 280nm. For an IgG using a 1cm light path an OD280 of 1.0 is equal to an antibody concentration 0.714mg/ml.

    When using Bradford type reagents, it is important to use an IgG standard curve. The absorbance generated by this type of reagent is dependent on the protein used. For example using a BSA standard curve to determine the protein concentration of an IgG solution will result in a two fold under estimate of the IgG concentration.

Properties

  • Storage instructionsStore at +4°C. Please refer to protocols.
  • Components 3 units
    Conjugation Buffer 1 unit
    Spin Cartridge/ Collecting tube assembly 3 units
  • Research areas

    Associated products

    Antibody Concentration Kit images

    Protocols

    References for Antibody Concentration Kit (ab102778)

    ab102778 has not yet been referenced specifically in any publications.

    Product Wall

    Thank you for your enquiry.
    The kit can be re-used up to 6 times. However, it can only be re-used with the same antibody. You could put wash buffer on its own through the column in between washes to clean it, but large particles will never get com...

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    Thank you for your call and your patience while I communicated with my colleague in the lab.

    In response to your inquiry, it turns out that both options (ab102784 Antibody Purification Kit (Protein A) or ab102778 Antibody Concentration Kit ) ...

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    The Antibody Concentration Kit method utilises a simple spin column to easily and quickly remove excess buffer from the antibody thereby providing a more concentrated antibody solution. Only molecules smaller than 10 kDa will be pass through the f...

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    Most benchtop microcentrifuges have a maximum spin of 14,000 - 15,000 RPM, and a maximum RCF of 18,000 - 21,000 x g. Anywhere in that range will be fine. You might want to test spinning just buffer without antibody, to see how long it takes to reduce 5...

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    The maximum volume of antibody that you can add to the spin cartridge is 0.5 mL. However, you are can use the spin cartridge multiple times to concentrate the 3 mL antibody sample.


    The laboratory responded that the filter in the antibody concentration kit ab102778 should work fine with a 30KD fragment. It is possible that the filter could be damaged. It would help if you could measure the OD of the flow through. If the flow ...

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    Thank you for contacting us. We do not currently sell ab15272 without sodium azide, but we may be able to custom formulate the antibody for bulk orders of more than 10 vials. For enquiries about a custom formulation, please contact mailto:sales@abcam.c...

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    Thank you for contacting us. We may be able to custom make this antibody in an azide-free form for bulk orders of 10 or more vials. If you are interested in a custom bulk purchase, please contact my colleagues at mailto:sales@abcam.com and they can hel...

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    Thank you for contacting us regarding ab58555.

    I have contacted the lab on your behalf, the antibody would be fine as well in a different buffer containing another stabilizing agent e.g. BSA. However this product is provided in PEG and suc...

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    Thank you for contacting Abcam regarding our EasyLink conjugation kits.


    I have reviewed the product specs of the antibody you are using and the buffer formulation is compatible with the HRP kit ab102891. As you mentioned, the only modific...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"