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Custom Conjugation Services

Need a conjugated antibody that isn't in the catalog? We can conjugate it for you.

  • Validated, optimized, and tested for stability with the same standards of the conjugated antibodies available in the catalog
  • Save time and avoid the pitfalls associated with do-it-yourself kits
  • Need to validate the conjugated antibody yourself? We also offer faster, lower-cost basic conjugation services

Contact us for more information.

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
Tier I Validation2 + Basic Service
Beads: Sepharose® and Magnetic Basic Conjugation + removal of free antibody
Tier I Validation3 + Basic Service
Enzymes: Horseradish peroxidase (HRP) Basic Conjugation + Purification
Tier I Validation2 + 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.
  • Flexibility: Tiered options range from basic antibody conjugation to full validation and stability testing of conjugates
  • Validation: Conjugates are tested in key applications using biologically relevant cell systems and controls, and verified for physical integrity using size exclusion chromatography
  • Optimization: Service options include choice of conjugation chemistries, removal of free dye and/or antibody purification, and identification of optimal degree of labeling (DOL) to ensure the best signal-to-noise ratio
  • Support: Application consultation and extensive technical support from the same scientists who produce and validate our commercially available antibody conjugates

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: loss of antibody during conjugation steps 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 (below).

Comparison of HRP-conjugated antibody in ELISA

18-WFO-15801 DIY ELISA comp data-v.3

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 from MCF7 cells untreated (dashed lines) or treated with human insulin-like growth factor I (solid lines) and the absorbance at 450 nm is shown (left). The signal-to-noise ratio was calculated at the 0.625 mg/ml lysate concentration (right).

Comparison of HRP-conjugated antibody in western blot

18-WFO-15801-Comparison of HRP-conjugated antibody in Western Blot-v.2

Comparison of HRP conjugation of GAPDH (D16H11) XP® Rabbit mAb using CST conjugation method or conjugated using a competitor’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 that signal specificity is better maintained with CST custom conjugation.

The following data illustrate performance of CST conjugated antibodies in a variety of applications. Note that the antibodies shown below are commercially available off-the-shelf; the same team of scientists performs conjugation and validation of both off-the-shelf and custom products.

Flow Cytometry

Flow cytometric analysis of human peripheral blood mononuclear cells

Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left column) or treated with cross-linked anti-CD3 plus anti-CD28 (10 µg/ml each, 15 min; right column), using Phospho-SLP-76 (Ser376) (D7S1K) XP® Rabbit mAb (PE Conjugate) #16318 (top row) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (bottom row), and co-stained with an antibody against CD3.

 

Flow cytometric analysis of PC-3 cells

Flow cytometric analysis of HeLa cells (blue) and KARPAS-299 cells (green) using TNFRSF8/CD30 (E7E4D) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 594 Conjugate) (dashed lines). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Western Blot

Western blot analysis of NF-κB Control Cell Extracts #9243 from HeLa cells

Western blot analysis of NF-κB Control Cell Extracts #9243 from HeLa cells, untreated (-) or treated with hTNF-α #8902, (20 ng/ml, 5 min., +), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Biotinylated) #4025.

Immunofluorescence

Confocal immunofluorescent analysis of mouse brain

Confocal immunofluorescent analysis of mouse brain using NeuN (D4G4O) XP® (Alexa Fluor® 488 Conjugate) #54761 (green). Actin filaments have been labeled using DyLight 554 Phalloidin #13054 (red).

Confocal immunofluorescent analysis of mouse primary bone marrow-derived macrophages or BMDM

Confocal immunofluorescent analysis of mouse primary bone marrow-derived macrophages (BMDMs) differentiated with mM-CSF (20 ng/mL, 8 days), and then either treated with LPS (50 ng/mL, overnight; left), or LPS (50 ng/mL, overnight) followed by Nigericin (10 uM, 2 hr; right), using ASC (D2W8U) Rabbit mAb (Mouse Specific) (Alexa® 488 Conjugate) #17507 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Note the translocation of ASC to inflammasomes following stimulation with LPS and Nigericin (white arrows).