Mouse Antibody Purification Kit (ab128745)

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.

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.

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.

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/Cytokeratin-8--18--19-antibody-2A4-ab41825.html (or use the following: https://www.abcam.com/Cytokeratin-8--18--19-antibody-2A4-ab41825.html).

https://www.abcam.com/EasyLink-Alkaline-phosphatase-Conjugation-Kit-1-x-100g-AP-ab102849.html (or use the following: https://www.abcam.com/EasyLink-Alkaline-phosphatase-Conjugation-Kit-1-x-100g-AP-ab102849.html).

https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html (or use the following: https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html).





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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:
https://www.abcam.com/index.html?pageconfig=resource&rid=13156#11

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