Technical Information | LYNX Rapid Conjugation Kits®

  1. Why are LYNX Rapid Conjugation Kits® superior to traditional kits and techniques?
  2. What can I label with LYNX Rapid Conjugation Kits®?
  3. What experimental conditions are needed?
  4. How long does antibody conjugation take?
  5. Do I need an extra step to separate out my labeled antibody?
  6. How do I prepare my conjugated antibody for use?
  7. Can I increase the amount of antibody I conjugate?
  8. How should I store my conjugated antibodies?
  9. Still have a question?

1) Why are LYNX Rapid Conjugation Kits® superior to traditional kits and techniques?

Faster, simpler procedure
LYNX Kits are easy to use — the simple one-step procedure needs only 30 seconds hands on time! Total conjugation time is either just 15mins with the LYNX Rapid Plus Conjugation Kits® or 3 hours with the LYNX Rapid Conjugation Kits®, but there is no problem with letting the reaction run overnight, if it is more convenient.

Antibodies ready-to-use — without further processing
Antibodies conjugated using LYNX Kits are ready for immediate use. There are no high molecular weight aggregates to be removed, and no desalting or dialysis steps are needed. This saves considerable time in comparison to standard conjugation techniques such as antibody thiolation or maleimide-activation of labels.

Consistent results
The simple one-step LYNX procedure involves no post-conjugation clean-up processes involving columns, dilution, concentration, and dialysis/desalting — the major sources of losses and batch-to-batch variation during conjugation. Eliminating these processes not only saves time, it also increases conjugation consistency and reduces sample loss. As the LYNX labeling reaction occurs in one vial, high yields of quality conjugated antibodies are the result — each and every time.

In addition, the easy reaction scale-up means that your results will be consistent no matter what amount of antibody you are labeling.

2) What can I label with LYNX Rapid Conjugation Kits®?

Antibody and protein labeling
LYNX Kits covalently conjugate label to amine groups to create a perfectly stable, high-quality conjugate that you can rely on. The primary application for LYNX Rapid Conjugation Kits® is to label purified antibodies, which have multiple amine groups. However, since most proteins contain many alpha-amino groups such as lysine, the kits can also be used to successfully label these molecules, opening up new avenues of research.

3) What experimental conditions are needed?

Antibody preparation for labeling
Purified antibodies are the ideal and recommended format for labeling with our LYNX kits. If antibodies are in ascites, serum or supernatant then all amine containing proteins in the solution will be labeled with the LYNX kit. Antibody preparations should not contain BSA as BSA will also be labeled with the LYNX kit.

Each LYNX kit has been optimized for labeling a particular concentration of antibody – please refer to the data sheet for further information about the optimal antibody concentration for the kit of your choice.

Buffers and additives for antibody labeling

Antibodies for conjugation should be prepared in phosphate*, HEPES, MES, MOPS or other amine-free buffers. Concentrations of up to 20 mM Tris buffer show almost no reduction in coupling efficiency.

The LYNX conjugation process is unaffected by additives such as sugars, salts, and chelating agents.

Buffer additives such as blockers (e.g. ethanolamine), nucleophiles (e.g. glycine), and thiols (e.g. DTT, 2-mercaptoethanol) may deactivate the conjugation process and should be avoided.

*BEWARE phosphate may inhibit alkaline phosphatase activity. Our alkaline phosphatase LYNX kits can be used with antibodies prepared in phosphate buffer as long as the buffer will be washed away during an immunoassay procedure. Otherwise we recommend using a non-phosphate containing buffer for labeling antibodies with the alkaline phosphatase LYNX kits.

Sodium azide
The presence of sodium azide in the antibody solution may slightly reduce the conjugation efficiency, and should be avoided if possible.

Sodium azide is not suitable for HRP conjugations as this compound is an irreversible inhibitor of HRP.

Physiological pH
LYNX Kits function at physiological pH, so antibodies are never exposed to the high pH required by some conjugation techniques.

4) How long does antibody conjugation take?

Actual hands-on time
It takes just 30 seconds of your time to conjugate an antibody with the simple LYNX one-step conjugation technique — regardless of sample size!

Total conjugation time
Conjugation is complete in just 15mins with the LYNX Rapid Plus Conjugation Kit® or 3 hours with the LYNX Rapid Conjugation Kit® and the labeled antibodies are ready for use after an additional 30 minutes. However, there is no negative effect from leaving the conjugation running overnight, if that is more convenient.

5) Do I need an extra step to separate out my labeled antibody?

Antibodies conjugated with LYNX Kits are suitable for use without the removal of any remaining free label. The simple one-step procedure needs no post-conjugation processing — eliminating one of the major sources of sample loss and batch-to-batch variation in traditional conjugation procedures.

Many applications using conjugated antibodies include a wash step, which will remove any remaining free label. If your application does not include a wash step then, if necessary, free label can be removed from the conjugated antibody by gel filtration. A sample volume of 5% or less of column volume should be used.

6) How do I prepare my conjugated antibody for use?

Desalting steps
The LYNX procedure deactivates all labeling reagents after conjugation is complete, so you can use the labeled antibodies directly for ELISA, immunohistochemistry, Western blotting, and most other applications without any desalting steps.

Buffers and additives for conjugated antibody
Conjugated antibodies can be diluted in any buffer compatible with both the label and antibody. For long term storage at 4°C, addition of antimicrobial agents, preservatives, or stabilizers (e.g sodium azide, BSA, glycerol, etc) is strongly recommended.

7) Can I increase the amount of antibody I conjugate?

Scalability
The LYNX Rapid Conjugation Kits® use a simple, consistent procedure for labeling both small and large quantities of antibody – the hands on time remains just 30 seconds! Antibody-conjugate preparations are consistent regardless of batch size, allowing you to initially conjugate microgram amounts of antibody for trial applications, and then scale up to milligram quantities when necessary.

8) How should I store my conjugated antibodies?

Conjugated antibodies can be stored in any buffer compatible with both the label and antibody. For long term storage at 4°C, addition of antimicrobial agents, preservatives, or stabilizers (e.g sodium azide, BSA, glycerol, etc) is strongly recommended. Other storage temperatures should be tested for suitability with small quantities of the conjugated antibody.

9) Still have a question?

If you have any other questions, contact our technical support team

 


LYNX Rapid Conjugation Kit® is a registered trademark of Bio-Rad AbD Serotec Ltd.