Custom AntibodyConjugation
Looking for a specific antibody conjugate? Cell Signaling Technology (CST) can conjugate it for you.
• Validated, optimized, and tested for stability to the same standards as the conjugated antibodies available in our catalog.
• Save time and avoid the pitfalls associated with do-it-yourself (DIY) kits.
• Need to validate the conjugated antibody yourself? We also offer basic conjugation services to match your timeline and budget needs.

Advantages of CST Custom Conjugation:

CST Custom Conjugates Deliver Superior Performance

Flexible: Tiered service offerings range from basic antibody conjugation
to full validation and stability testing of conjugates.

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.

Application-Validated Antibody Conjugates

59.2

DIY Conjugation Kit

17.6

EventsValidated: Conjugates are tested in key applications using biologically
relevant cell systems and controls.

Signal-to-NoiseCST Custom Conjugation

CST CUSTOM CONJUGATIONDIY CONJUGATION KIT
Helios PE Conjugate
DIY Conjugation Kit compared to CST Custom Conjugation: Comparison of PE conjugation of Helios (D8W4X) XP® Rabbit mAbby flow cytometry, using CST custom conjugation method (blue) or another company’s DIY conjugation kit (green), on both
RL-7 (dashed lines), a low-expressing cell line, and Jurkat (solid lines), a high-expressing cell line. A comparison of thecalculated signal-to-noise ratio is also shown. Avoid the inefficiencies caused by DIY conjugation such as suboptimallabeling, interference with specificity, and/or destabilization/degradation that can negatively affect antibody performanceand decrease yields.

The following data illustrate the performance of Cell Signaling Technology (CST) ready-made conjugated antibodies in different applications.
These are representative of the quality and validation of custom conjugates from CST.
Flow Cytometry

Immunofluorescence
Anti-CD3/Anti-CD28 Treated
CD3 (UCHT1) (APC Conjugate)

CD3 (UCHT1) (APC Conjugate)

Untreated

Phospho-SLP-76 (Ser 376)
(Alexa Fluor® 488 Conjugate)
CD3 (UCHT1) (APC Conjugate)

CD3 (UCHT1) (APC Conjugate)

Phospho-SLP-76 (Ser 376)
(Alexa Fluor® 488 Conjugate)

Rabbit IgG Isotype Control(Alexa Fluor® 488 Conjugate)

Rabbit IgG Isotype Control(Alexa Fluor® 488 Conjugate)

Phospho-SLP-76 (Ser376) (E3G9U) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #47876: Flow cytometric analysis
of human PBMCs, untreated (left column) or treated with cross-linked anti-CD3 plus anti-CD28 (right column), using
Phospho-SLP-76 (Ser376) (E3G9U) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) (top row) or concentration-matched
Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (bottom row). Co-stained with CD3 (UCHT1)
Mouse mAb (APC Conjugate) #19881.

TIPE2 (E5Q2S) Rabbit mAb #53842: Confocal IF analysis of mouse colon using TIPE2 (E5Q2S) Rabbit mAb (green). After blocking
free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissues were then labeledusing Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #70034 (cyan pseudocolor) and CD45 (D3F8Q) Rabbit mAb (Alexa Fluor® 647 Conjugate) #81143 (red). Samples were mounted in ProLong® Gold
Antifade Reagent with DAPI #8961 (blue).

For Research Use Only. Not For Use In Diagnostic Procedures.

www.cst-science.com/customconjugation

Custom Antibody Conjugation

S/N Ratio

Degree of Labeling (DOL) to Ensure Specific Signal Intensity

Conjugation Services Available:

45Conjugation Type

Service Tier

Service Includes

40

Fluorophores Type I:

BasicConjugation + removal

35

Alexa Fluor , Cyanine

30(CyDye ) and Pacific Blue

25dyesof free dye

®

®

™

Tier I+ Basic Service
Tier II

20

DOL 4.28

105

Tier III

Tier II Service

DOL 5.51

Fluorophores Type II:

BasicConjugation + purification

DOL 7.27

Phycoerythrin (PE),Tier I

Validation1 + Basic Service

PE-CyDye® tandems

Tier II

Stability testing +0Concentration (µg/ml)

DOL service option for fluorophores: Conjugation to certain fluorescent dyes (Alexa Fluor® and Pacific Blue™)is optimized by degree of labeling (DOL) testing to identify the optimal antibody:dye molecular ration, resulting
in conjugates with maximum specific signal intensity.

Size Exclusion to Isolate Oligo-antibody Conjugate
Unconjugated Oligo

Tier I Service(e.g. PE-Cy 7), and®

allophycocyanin (APC)

Haptens: Biotin,

Basic

Unconjugated Antibody

DNP, and Dansyl

Tier I

Validation2 + Basic Service

Beads: Sepharose®,

Basic

Conjugation + removalof free antibody

Tier I

Validation3 + Basic Service

Basic

Conjugation + purification

peroxidase (HRP)

Tier I

Validation2 + Basic Service

Oligonucleotides4

BasicConjugation + purification

Tier I

Biophysical Characterization

Enzymes: Horseradish

Oligo-Antibody Conjugate

Conjugation + removalof free biotin

Digoxigenin (DIG),

Magnetic, Agarose
Absorbance (mAU)

Validation1 + Tier I Service
Stability testing +DOL 2.34

15

Degree of labeling (DOL)+ Basic Service
Time (minutes)
Final Conjugate Isolated from Unconjugated starting materials: Size Exclusion Chromatogram comparing purified
Oligo-Antibody Conjugate (green peak) to unconjugated Antibody (blue line) and unconjugated Oligo (red line).
CST uses Size exclusion chromatography to separate out free materials and deliver only the purified Oligo-Antibody
conjugate product to the customer, ensuring a higher quality reagent.

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

Interested in a custom conjugation or a specific conjugation type not listed above?
Contact us: cst-science.com/conjugationrequest

Oligonucleotide Conjugation Services

20-ETC-49293-FLY1-E1-JAN23• 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.

©2
 023 Cell Signaling Technology, Inc. Cell Signaling Technology, CST, and XP are trademarks of Cell Signaling Technology, Inc. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

For Research Use Only. Not For Use In Diagnostic Procedures.

www.cst-science.com/customconjugation