• Product name
    Mouse Antibody Purification Kit
    See all Antibody Purification kits
  • Species reactivity
    Reacts with: Mouse
  • Product overview

    Commercially available antibodies often contain substances (e.g. BSA, glycine, tris, azide) that interfere in labeling reactions with enzymes or fluorophores. ab128745 quickly removes these contaminants. It can also be used to purify antibodies from crude samples such as ascites fluid or immune serum. The mouse antibody to be purified or cleaned up ideally is in a volume of 100µl to 0.5ml. 20µg to 150µg of antibody can be purified in each run.

    The method involves capture of the antibody on a mouse resin and the removal of unwanted substances by a simple wash procedure, which is carried out in a standard microfuge. The purified product is then eluted and neutralized. Note: The Mouse Antibody Purification Kit is not suitable for use with antibodies from other species. It should be noted that the binding strength of Bovine IgG to the Mouse resin is negligable.

    The components of ab128745 are fully compatible with our Conjugation kits however they are not compatible with our GOLD mouse Antibody conjugation kits. To purify antibodies for use with our GOLD conjugation kits, please use our GOLD mouse anibody purification kit (ab204910).


    ab128745 is not suitable for use with antibodies from species other than mouse.

  • Notes



    1. Reconstitution of Mouse Resin

    Add 0.3ml of wash buffer to each vial of Mouse Resin, mix by inversion for a few seconds and transfer to the spin cartridge. Spin for 30 seconds in a microfuge.


    2. Incubation of Sample with Resin

    To the antibody, add an appropriate amount of 10x Binding Buffer. For example, if the sample volume is 200µl, add 20µl of Binding Buffer. Pipette the sample into the spin cartridge and cap the tube. Incubate for a minimum of 2 hours with agitation or periodic shaking. Alternatively, incubate overnight at 4°C or at room temperature.

    Note: The volume of antibody to be purified or cleaned up ideally should be 0.1-0.5ml, though larger volumes may be processed by first incubating the antibody sample with the mouse specific resin in a larger vessel (e.g. 2ml tube) prior to transferring to the spin cartridge.

    3. Wash Procedure

    Microfuge the spin cartridge assembly for 30 seconds to remove most of the non-bound protein. Add 0.5ml of wash buffer and spin again. Repeat the wash procedure three times.

    Note: Save the non-bound and wash fractions by transferring the material from the collecting tube after each spin to a set of eppendorfs (not supplied). Do not use the four collecting tubes supplied with the kit, as these have an extended hinge to accommodate the spin cartridge, and are required for the elution step.

    4. Elution

    1. Transfer the cartridge to a clean collecting tube. Add 100µl of elution buffer and incubate for 2 min at room temperature with gentle agitation. Microfuge for 30 seconds. Remove the collecting tube and add 25µl neutralizer to the tube.
    • Place the cartridge in a new collecting tube and add a further 100µl of elution buffer to the mouse resin. Incubate for 2 min at room temperature with gentle agitation. Spin and collect and neutralize as before.


    • Repeat the elution procedure until all four clean collecting cups have been used. The protein normally elutes in tubes 1 and 2 but you should confirm this using a test for protein (see below) before pooling any of the tubes.


    • Pool the tubes with most protein (normally two tubes; if more than two tubes are strongly positive it is possible that you have used too much sample in your protein assay). However, if your application does not require a high concentration of antibody you may choose to pool all tubes that contain protein, regardless of concentration.


    Storage of Antibody

    Store at 4°C. Other storage conditions (e.g. frozen at -80°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 280 nm.

    When other methods are used such as BCA or Bradford protein assays, determinations should be performed before the addition of the neutralization buffer. The neutralization buffer contains components that can interfere with these reagents. The neutralization buffer should be added to the sample as soon as possible as the low pH of the elution buffer can denature the antibody.

    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.

  • Tested applications
    Suitable for: Purificationmore details


  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components Identifier 1 tests 3 tests
    10x Binding Buffer 1 vial 1 vial
    Collecting Tubes 4 units 12 units
    Elution Buffer 1 vial 1 vial
    Mouse Resin White cap 1 vial 3 vials
    Neutralizer Buffer 1 vial 1 vial
    Spin Cartridges/ Collection tubes assemblies 1 vial 3 vials
    Wash Buffer 1 vial 1 vial
  • Research areas


Our Abpromise guarantee covers the use of ab128745 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Purification Use at an assay dependent concentration.



ab128745 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A


The Mouse Antibody Purification Kit was developed to purify mouse IgGs and is suitable to purify any mouse IgG. This product was never tested on IgAs, although we do not think it will be as efficient as it is for IgG purification.

Read More


Based on the description of the thicker, coagulated substance it sounds like the antibody precipitated in the low pH elution buffer. If this is the case then the precipitated protein will not go through the spin column, hence the absence of an absorbance at 280nm in the eluted fractions.
Would you mind sharing the information about the specific antibody that you were trying to purify? Is there any information on how it reacts to different pH levels? With additional information we may be able to come up with an alternative approach.

Read More


The antibody purification kits that we carry, such as ab128745 and ab128747, will only recognize and purify IgG. For other classes such as IgA, IgD, and IgM, we have class-specific antibodies, for instance ab124716, which could be used to immunprecipitate those classes of immunoglobulin. Please let me know if you have any questions.

Read More


Vielen Dank für Ihren Anruf.

Leider sind 0,5% BSA das Maximum, dass das Labeling-Kit toleriert. Daher wäre entweder eine IP notwendig, mit der Sie den Antikörper von BSA aufreinigen notwendig, oder aber Sie benutzen ein weiteres Kit priori dem eigentlichen Labeling Kit.

Zusammenfassend lässt sich sagen dass Sie die folgenden Produkte von uns zur Konjugation erwerben können:

https://www.abcam.com/index.html?datasheet=41825 (or use the following: https://www.abcam.com/index.html?datasheet=41825).

https://www.abcam.com/index.html?datasheet=102849 (or use the following: https://www.abcam.com/index.html?datasheet=102849).

https://www.abcam.com/index.html?datasheet=128745 (or use the following: https://www.abcam.com/index.html?datasheet=128745).

Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!

Read More


As we discussed, our studies have found little effect on the conjugation chemistry with up to 0.05% BSA and up to 0.1% glycine in the antibody storage buffer. Thus, I think you do not have to worry about those components. However, as you know the 0.01% sodium azide will inhibit HRP activity, and as you mentioned the 50% glycerol is making it hard to remove the sodium azide.

Antibody purification kit if you are going to stay with ab76424:
Protein A for rabbit antibodies ab102784: https://www.abcam.com/Antibody-Purification-Kit-Protein-A-ab102784.html
Protein G for other species ab 128747: https://www.abcam.com/Antibody-Purification-Kit-Protein-G-ab128747.html
For mouse antibodies ab128745: https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html

Previous customers have contacted us with issues purifying antibodies with high concentrations of glycerol, and we found the following solution proved successful:
" When using the antibody purification kit the presence of glycerol in the antibody buffer will make it more viscous. This means that it will require more G force to pass through the spin column. It is possible to buffer exchange the antibody it just means the customer will have to spin for much longer. Alternatively, a simpler solution would be to dilute the antibody. This would reduce the glycerol’s concentration, which in turn will make the buffer less viscous and easier to spin out. You can then use the same spin column to concentrate the antibody back to its original concentration and volume, after having performed the buffer exchange."

Additionally, we have our EasyLink conjugation kits, including HRP conjugation, listed through the link below. Our EasyLink conjugation kits are capable of fully successful conjugation with glycerol concentrations of up to 50%. We are currently running a promotion for an extra 20% off our EasyLink kits:
EasyLink Kits: https://www.abcam.com/index.html?pageconfig=resource&rid=13148#enzymatic
Easylink Promotion:https://www.abcam.com/index.html?pageconfig=resource&rid=15389&source=pagetrap&viapagetrap=easylink123
Below is a list of compatible and incompatible buffers with our EasyLink conjugation reactions:

Read More


Thank you for your reply.

You are spot on about using the Mouse antibody purification kit, ab128745, to remove the BSA, sodium azide, glycine, tris, and other molecules that may be present in a volume of 100-500ul. The kit is compatible with ab128745 and other TCS antibodies. ab128749 is listed as "TCS" in its title for purifying antibody from large, non-purified, TCS volumes.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

For licensing inquiries, please contact partnerships@abcam.com

Sign up