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AqT™ Biotin Active Ester Antibody Labeling Kit (T2A15)

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Price:
$149.00
SKU:
CM86140.1
Brand:
Availability:
In stock
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Calculated at checkout
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SDS USER MANUAL CM86140

AqT™ Biotin Active Ester Antibody Labeling kit (T2A15) is a proprietary biotinylation kit developed at CellMosaic for obtaining high quality and high loading of biotinylated antibody with minimal or no aggregation. The kit uses a biotin modified with a super-hydrophilic, water-soluble, and charge neutral AqueaTether (AqT) T2A15 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. AqT™ T2A15 linker composed of two threitols and 15 atom lengths. This atom length is equivalent to PEG4 (16 atom lengths) and, based on our experience, can largely keep the bivalent molecule function independent. The conjugates we have tried with this atom length include antibody oligo, streptavidin oligo, and HRP oligo, among others.

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. A quick 10-minute biotin assay (protocol and reagent) is also included in the kit to obtain the biotinylation level immediately after labeling. 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. 

Catalog Number

Number of Reactions in Kit

Antibody Amount per Reaction

CM86140.1x1

1

10 μg

CM86140.1x3

3

CM86140.2x1

1

50 μg

CM86140.2x3

3

CM86140.3x1

1

100 μg

CM86140.3x3

3

CM86140.4x1

1

1 mg

CM86140.4x3

3

CM86140.5x1

1

3 mg

CM86140.5x3

3

 


CM86140 and Its Product Compared to Other Commercial Antibody Biotinylation Kits

 

 

AqT™ Biotin Labeling Kit

Other Commercial Kits

Biotinylated Product Features

 

Chemical structure

Fully Disclosed

·  Complete knowledge of your product

Varies by vendor, mostly not disclosed.

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

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

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

Labeling buffer

100% aqueous buffer

Some percentage of organic solvent needed, such as DMSO

Biotinylation level

Predictable

·  Tunable loading, from low to very high, per your need

Unpredictable

Usually target low labeling to avoid aggregation (difficult to tune)

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

Purification method & components

Included

Product is free of any unreacted AqT™ biotin after purification.

Varies by vendor, usually not included

Biotin Assay

Included

A quick 10-minute biotin assay protocol with reagents and buffer

Not included


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-biotinylation-chemistry.png


Overview of the workflow for preparing AqT™ antibody-biotin conjugates

cm86140-protocol.png

 


Comparing the hydrophilicity of AqT biotin-T2A15 acid with biotin and biotin-PEG4 acid (C18 HPLC Profile)

overall-aqt-biotin-full-length.png


Comparing the hydrophilicity and heterogenicity of AqT biotin-T2A15 labeled antibody with unmodified biotin labeled antibody (HIC HPLC Profile)

 aqt-technology-page-figure5.png


Comparing the apparent MW (hydrodynamic volume) of AqT biotin-T2A15 labeled antibody with unmodified biotin labeled antibody (SEC HPLC Profile)

aqt-technology-page-figure6.png

Other Details

Scale of the Reaction:
10 ug-3 mg
Antibody Labeling Sites:
Surface amines

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