Antibody purification and concentration kits
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Select the right kit for your antibody purification and concentration needs with our guide
Overview
Select the right kit for your antibody purification and concentration
Our easy-to-use flow diagrams will help you choose the best antibody purification kit or antibody concentration kit for your needs.
| Purification selection - kits | Purification selection - nanoparticles |
| Select a kit that is compatible with our Lightning-Link®, Oligonucleotide, and metal conjugation kits | Select a kit that is compatible with our nanoparticle conjugation kits |
Antibody purification kits
Many commercially available and in-house manufactured antibodies often contain substances that may interfere with applications such as antibody conjugation, making purification a key consideration. Our antibody purification kits are suitable for purifying antibodies from serum, tissue culture supernatant (TCS), and ascites fluid. The resulting purified antibodies are fully compatible with our conjugation kits.
Purification of antibodies from unwanted buffer constituents (eg, BSA)
Antibodies are often stored in buffers that contain substances other than the antibody itself. Our antibody purification kits highlighted in Table 1 are designed to remove protein additives, leaving the antibody in a format suitable for transfer to a conjugation-friendly buffer.
Table 1.
| Product name | Use | Conjugation kit compatibility |
| Antibody Purification Kit (Protein A) (ab102784) | Suitable for purifying IgG from multiple species Removes contaminants including BSA, glycine, tris, and azide Purifies antibodies from crude samples such as ascites fluid or immune serum | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Antibody Purification Kit (Protein G) (ab128747) | Suitable for purifying IgG from multiple species Removes contaminants including BSA, glycine, tris, and azide Purifies antibodies from crude samples such as ascites fluid or immune serum | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Mouse Antibody Purification Kit (ab128745) | Suitable for purifying mouse IgG only Removes contaminants including BSA, glycine, tris, and azide Purifies antibodies from crude samples such as ascites fluid or immune serum | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| BSA Removal Kit (ab173231) | Suitable for purifying any antibody sub-type and species Removes BSA, glycine, tris, and azide | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Antibody Purification Kit – Nanoparticles (ab204909) | Suitable for purifying IgG from multiple species Removes contaminants including BSA, glycine, tris, and azide Purifies antibodies from crude samples such as ascites fluid or immune serum | Nanoparticle conjugation kits |
| Mouse Antibody Purification Kit – Nanoparticles (ab204910) | Suitable for purifying mouse IgG only Removes contaminants including BSA, glycine, tris, and azide Purifies antibodies from crude samples such as ascites fluid or immune serum | Nanoparticle conjugation kits |
| BSA Removal Kit – Nanoparticles (ab204912) | Suitable for purifying any antibody sub-type and species Removes BSA glycine, tris, and azide | Nanoparticle conjugation kits |
The method involves capturing the antibody on agarose beads coupled to proteins A or G, or resin that has affinity and specificity for mouse IgG for the mouse antibody purification kits. After incubation of the antibody with the beads or resin, a simple wash procedure removes any unwanted substances. The purified antibody is then eluted and neutralized. This method cannot be used with samples containing dilute antibody in large volumes, eg, tissue culture supernatant.
Purification of antibodies from TCS
TCS contains bovine IgG molecules from fetal bovine serum (FBS), which supplements cell growth media. These molecules need to be removed before carrying out conjugation reactions because they can compete with the antibody for labeling.
Table 2.
| Product name | Use | Conjugation kit compatibility |
| TCS Antibody Purification Kit (ab109207) | Suitable for purifying IgG from most species, not suitable for purifying goat IgG | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Mouse TCS Antibody Purification Kit (ab128749) | Suitable for purifying mouse IgG only | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Rat TCS Antibody Purification Kit (ab270562) | Suitable for purifying rat IgG only | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| TCS Antibody Purification Kit – Nanoparticles (ab270554) | Suitable for purifying IgG fractions | Nanoparticle conjugation kits |
The purification kits listed in Table 2 use protein A or resins with a high affinity and specificity for mouse or rat IgG molecules coupled to agarose beads. The resin can selectively capture the antibody without co-purifying bovine IgG as its binding strength for bovine IgG molecules is minimal.
Following incubation, the resin is loaded into a column, and a simple wash procedure removes unwanted substances. The purified antibody is then eluted and neutralized.
Purification of antibodies from serum or ascites fluid
Our antibody serum purification kits (Table 3) are prepared by coupling highly purified protein A or G to agarose beads to purify numerous components from antibodies generated from ascites fluid or serum.
Table 3.
| Product name | Use | Conjugation kit compatibility |
| Serum Antibody Purification Kit (Protein A) (ab109209) | Suitable for purifying IgG fractions from serum and ascites fluid Not suitable for purifying goat IgG* | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Serum Antibody Purification Kit (Protein G) (ab128751) | Suitable for purifying IgG fractions from serum and ascites fluid | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Antibody Purification Kit - Nanoparticles (ab204909) | Purifies antibodies from crude samples such as ascites fluid or immune serum | Nanoparticle conjugation kits |
* Please refer to the protocol booklet for more information about suitable and unsuitable species antibodies.
The antibody is captured on the agarose beads and incubated with the resin. Following the incubation period, the resin is loaded into a column, and a simple wash procedure removes unwanted substances. The purified antibody is then eluted and neutralized.
The magnetic antibody purification system is a protein A-based antibody purification system with a unique set of elution and neutralization buffers used for the concentration of antibodies. It is also able to purify antibodies at a small scale with high recoveries.
The antibody binds to protein A magnetic beads, which are subsequently captured with a magnetic stand to purify the antibody.
Table 4.
| Product name | Use | Conjugation kit compatibility |
| Magnetic Antibody Purification System (ab270532) | Antibody concentration Suitable for purifying very low amounts of antibody | Compatible with all conjugation kits |
Antibody concentration and clean-up kits
Our antibody concentration and clean-up kits (Table 5) enable the quick and easy concentration of antibodies and proteins using a spin column.
Table 5.
| Product name | Use** | Conjugation kit compatibility |
| Antibody Concentration And Clean-Up Kit (ab102778) | Suitable for the concentration of antibodies and proteins Remove small additives such as azide, glycine, and Tris | Lightning-Link® conjugation kits Oligonucleotide conjugation kits |
| Antibody Concentration And Clean-Up Kit – Nanoparticles (ab204911) | Suitable for the concentration of antibodies and proteins Remove small additives such as salt, citrate, and azide | Gold and Magnetic Conjugation Kits |
| Antibody Concentration and Clean Up Kit for Latex & Europium (ab269965) | Suitable for the concentration of antibodies and proteins Remove small additives such as salt, citrate, and azide | Latex and Europium conjugation kits |
** It is essential to note that any buffer components with a molecular weight greater than 10 kDa, such as BSA, will also become concentrated during this process. If the additive concentration is incompatible for conjugation following concentration and resuspension, the appropriate purification kit should be used.
These kits can also be used to remove any undesirable low molecular weight additives such as Tris, glycine, and sodium azide from the antibody storage buffer that passes through a filter, trapping the antibody, which is then recovered in a conjugation-friendly buffer. Removing sodium azide is particularly important if conjugating your antibody Horseradish Peroxidase (HRP) because sodium azide rapidly reduces HRP’s enzyme activity.