Custom Antibody Conjugation

Need a conjugated antibody that you don’t see in the Cell Signaling Technology^® (CST^®) catalog? Save time and avoid pitfalls, like the low yields associated with do-it-yourself (DIY) kits. We can conjugate it for you.

Advantages of CST Custom Conjugation:

Flexible: Tiered service offerings range from basic antibody conjugation to full validation and stability testing of conjugates.
Validated: Conjugates are tested in key applications using biologically relevant cell systems and controls, and verified for physical integrity using size exclusion chromatography.
Optimized: Flexible conjugation chemistry, free dye and/or antibody purification, and identification of optimal degree of labeling (DOL) are utilized to ensure the best signal-to-noise ratio.
Support: Receive consultation and technical support from the conjugation experts who produce our ready-made conjugates.

REQUEST YOUR CUSTOM ANTIBODY CONJUGATION

Conjugation Services available:

Conjugation Type: Examples	Service Tier	Service Includes
Fluorophores Type I: Alexa Fluor^®, Cyanine (CyDye^®) and Pacific Blue^™ dyes	Basic	Conjugation + removal of free dye
	Tier I	Degree of labeling (DOL) + Basic Service
	Tier II	Validation¹ + Tier I Service
	Tier III	Stability testing + Tier II Service
Fluorophores Type II: Phycoerythrin (PE), PE-CyDye^® tandems (e.g. PE-Cy^®7), and allophycocyanin (APC)	Basic	Conjugation + purification
	Tier I	Validation¹ + Basic Service
	Tier II	Stability testing + Tier I Service
Haptens: Biotin, Digoxigenin (DIG), DNP, and Dansyl	Basic	Conjugation + removal of free biotin
Haptens: Biotin, Digoxigenin (DIG), DNP, and Dansyl	Tier I	Validation² + Basic Service
Beads: Sepharose^® and Magnetic	Basic	Conjugation + removal of free antibody
Beads: Sepharose^® and Magnetic	Tier I	Validation³ + Basic Service
Enzymes: Horseradish peroxidase (HRP)	Basic	Conjugation + Purification
Enzymes: Horseradish peroxidase (HRP)	Tier I	Validation² + Basic Service
Oligonucleotide ⁴	Basic	Conjugation + Purification
Oligonucleotide ⁴	Tier I	Biophysical Characterization + Basic Service

¹ Standard validation application method is Flow Cytometry. Alternatives may increase timeline and pricing.
² Standard validation application method is Western Blot. Alternatives may increase timeline and pricing.
³ Standard validation application method is IP. Alternatives may increase timeline and pricing.
⁴ Oligonucleotide conjugation offering may be limited for certain IP protected technologies.

Oligonucleotide Conjugation Services

Biophysical characterization checks integrity, size, and confirms no aggregation, free antibody or free oligo, and is typically performed by analytical chromatogram.
Basic Certificate of Analysis (COA) includes Oligo sequence, antibody concentration, and DOL information. Tier I COA also includes biophysical characterization results.
DOL offering allows one to analyze individual fractions or multifraction pools to select the best product.

CST Custom Conjugation vs Do-It-Yourself (DIY) Conjugation Kit

DIY conjugation kits look good on paper, but are you getting your money’s worth? Inefficiencies caused by suboptimal labeling, interference with specificity, and/or destabilization/degradation can negatively affect performance. Another drawback is the loss of antibody during conjugation steps, which can mean finishing with significantly less functional antibody than you started with.

With Custom Conjugation Services from Cell Signaling Technology (CST), you can avoid these time-consuming pitfalls and get the full amount of antibody you pay for.

Comparison of PE-conjugated Antibody in Flow Cytometry

18-WFO-15801 Comparison of PE Conjugated Antibody in Flow Fever Chart

Comparison of PE conjugation of Helios (D8W4X) XP^® Rabbit mAb using CST custom conjugation method (blue) or conjugated using a competitor’s DIY PE conjugation kit (green). Flow cytometeric analysis of RL-7 (dashed lines), a low-expressing cell line, and Jurkat (solid lines), a high-expressing cell line. The calculated signal-to-noise ratio is shown (above).

Comparison of HRP-conjugated Antibody in ELISA

Comparison of HRP conjugation of Phospho-Akt (Ser473) (193H12) Rabbit mAb conjugated using CST custom conjugation method (blue) or conjugated using a competitor’s DIY HRP conjugation kit (green), performed by ELISA assay. The relationship between the protein concentration of lysate (x-axis) from MCF7 cells untreated (dashed lines) or treated with human IGF I (solid lines) and the absorbance at 450 nm is shown (y-axis). The signal-to-noise ratio was calculated at the 0.625 mg/ml lysate concentration (in associated table).

Comparison of HRP-conjugated Antibody in Western Blot

Comparison of HRP conjugation of GAPDH (D16H11) XP^® Rabbit mAb using CST conjugation method or conjugated using a another company’s DIY HRP conjugation kit. Western blot analysis of extracts from C6 and THP-1 cells were performed with concentration-matched antibody conjugates. These data demonstrate the increased signal specificity observed when using CST custom conjugation versus a DIY HRP conjugation kit.

Regarding Alexa Fluor conjugation, this product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or [email protected]. Alexa Fluor^® is a registered trademark of Life Technologies Corporation. Cy, Cy7, CyDye. Pacific Blue is a registered trademark of Molecular Probes, Inc. All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.