Product Description
AqT® Biotin Active Ester Antibody Labeling kit is a proprietary biotinylation kit developed at CellMosaic® for obtaining high quality and high loading of biotinylated antibody with minimal or no aggregation.
| Download Documents | |
| SDS | USER MANUAL CM86140 |
The kit uses a biotin modified with a super-hydrophilic, water-soluble, and charge neutral AqueaTether® (AqT®) linker to improve its water solubility and decrease its non-specific interaction with other proteins or plastic surfaces. Biotinylated antibodies with AqT® linker can largely retain its native properties and keep the bivalent molecule function independent.
For a detailed description of the AqT® technologies, please see our technology section.
The CM86140 kit is designed to biotinylate any antibody via surface amines of the antibody. The user supplies the antibody. Using the kit components, the user biotinylates their antibody in one simple step, followed by filtration-type purification to remove any unreacted AqT®-biotin, exchanging it with PBS or other buffer for downstream application. The biotinylation level can easily be tuned by increasing or decreasing the reaction temperature. The hands-on time is less than 30 minutes, and the whole preparation time varies from 3 h to overnight depending on the reaction time.
The kit has multiple configurations for labeling various amounts of antibody (10 μg to 3 mg) with either one (x1) or three (x3) antibody samples.
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Catalog Number |
Number of Reactions in Kit |
Antibody Amount per Reaction |
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CM86140.1x1 |
1 |
10 μg |
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CM86140.1x3 |
3 |
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CM86140.2x1 |
1 |
50 μg |
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CM86140.2x3 |
3 |
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CM86140.3x1 |
1 |
100 μg |
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CM86140.3x3 |
3 |
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CM86140.4x1 |
1 |
1 mg |
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CM86140.4x3 |
3 |
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CM86140.5x1 |
1 |
3 mg |
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CM86140.5x3 |
3 |
Requirements for antibody:
- MW: ≥ 100 KDa (if MW is lower, please use an alternative kit)
- Purity: preferably > 90% pure by gel electrophoresis
- Amount: 10 μg to 3 mg protein content as measured by UV
Labeling Chemistry
CM86140 and Its Product Compared to Other Commercial Antibody Biotinylation Kits
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AqT® Biotin Labeling Kit |
Other Commercial Kits |
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Biotinylated Product Features
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Chemical structure |
Fully Disclosed · Complete knowledge of your product |
Varies by vendor, mostly not disclosed. |
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Properties |
Retain antibody properties · No changes in hydrophobicity · No increase of apparent MW (hydrodynamic volume) · No increase in aggregation · Minimal heterogeneity after labeling |
Change antibody properties · Increased hydrophobicity (non-specific interaction) · Increased apparent MW (hydrodynamic volume) · May increase aggregation · Highly heterogeneous product |
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Stability |
Stable even with high biotin loading Can be easily stored frozen and in lyophilized form without any stabilizer |
Decreased stability High biotin loading may cause aggregation and precipitation |
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Kit Features |
Reagent |
Proprietary water-soluble pre-activated biotin with 15-atom spacer |
Biotin (zero length) or biotin with hydrophobic ethylene- or polyethylene glycol-type linker |
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Labeling buffer |
100% aqueous buffer |
Some percentage of organic solvent needed, such as DMSO |
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Biotinylation level |
Predictable · Tunable loading, from low to very high, per your need |
Unpredictable Usually target low labeling to avoid aggregation (difficult to tune) |
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Usability |
Easy to use · One step labeling · <30 minutes hands-on time |
Comparable quick labeling may use EDC chemistry with antibody crosslinking to each other causing large aggregates |
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Purification method & components |
Included Product is free of any unreacted AqT™ biotin after purification. |
Varies by vendor, usually not included |
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Biotin Assay |
Included A quick 10-minute biotin assay protocol with reagents and buffer |
Not included |
Overview of the workflow for preparing AqT® antibody-biotin conjugates
Comparing the hydrophilicity and heterogenicity of AqT® biotin labeled antibody with unmodified biotin labeled antibody (HIC HPLC Profile)
Comparing the apparent MW (hydrodynamic volume) of AqT® biotin labeled antibody with unmodified biotin labeled antibody (SEC HPLC Profile)
Frequently Asked Questions:
The list below includes FAQs that are specific to this product.
Question: I need to mono-biotinylate my 15 KDa nanobodies with 3 to 4 lysine residues. Can CM86140.1 be used or is there some adjustment for your current method to make this mono-biotinylation happen?
Answer: CM86140 is designed for labeling antibody but can also be used for labeling other proteins with some adjustment (both hardware and user manual). The loading can be adjusted by changing the amount of biotinylation reagent. However, keep in mind that, unless you have tried to see the reactivities of the lysine residues, it is difficult to know what your biotinylation level might be. The best adjustment can be made by using the data obtained from the first labeling experiment. I don’t know how critical you want your 1:1 loading. Since you have 3 to 4 lysine residues, your conjugate will be a mixture of unlabeled, 1:1 labeled, 1:2 labeled, and so on. If you target 1:1 labeling, you will have some unlabeled nanobody, which can be removed by HPLC. The problem is how to find your labeling ratio. If you aim for labeling 10 to 50 micrograms, the included biotinylation assay may not be very accurate due to the low amount and loading. You may have to perform MS analysis to check the loading.
If you can’t find the answer you’re looking for or need information on general topics, please visit the main Frequently Asked Questions (FAQs) section.
