Thunder-Link® PLUS Oligo Conjugation System

Thunder-Link® PLUS enables simple and rapid conjugation of antibodies to oligonucleotides, with high recovery of materials and a superior clean-up procedure. The kit is quick and simple to use, overcoming time consuming and lengthy protocols associated with standard conjugation methods.

Thunder-Link®  PLUS Features and Benefits

  • Quick and easy to use – Save time, no specialist knowledge required
  • High levels of antibody and oligo recovery – Save precious reagents
  • Use your own oligo and antibody, at your desired ratio – Flexible
  • Freeze dried – Ships at ambient temperature, long shelf-life
  • Stringently QC tested – Consistent high quality, excellent batch-to-batch reproducibility
  • Unidirectional chemistry – No risk of cross-linking
  • Covalent bond – Highly stable conjugates
  • Suitable for single-stranded oligos of 10-120 bases, double-stranded oligos up to 80 base pairs – Covers the majority of sequences
  • Linking chemistry works at both 5’ and 3’ end – Provides ability to combine with other modifications
  • Post-conjugation clean-up step – No interference from unbound oligo
  • Positive control antibody and oligo included – Enables confirmation of protocol success
  • Wide range of target proteins – Also applicable to antibody fragments and small proteins

Main applications: immuno-PCR, Proximity Ligation Assay, Electrochemical Proximity Assay

Advantages of Thunder-Link® PLUS:

  • Only 30 minutes antibody and oligo activation
  • Fast oligo conjugation – now only 1 hour!
  • High antibody and oligo recovery
  • Robust and flexible clean-up procedure (now works with antibody fragments and other proteins)

Thunder-Link® PLUS Kit Components

  1. Thunder-Link® PLUS Oligo Activation reagent
  2. Thunder-Link® PLUS Antibody Activation Reagent
  3. Freeze dried control oligo
  4. Freeze dried control antibody
  5. Separating columns
  6. Thunder-Link® PLUS Wash buffer
  7. Thunder-Link® PLUS Conjugate Clean Up Reagent
  8. Thunder-Link® PLUS Antibody Suspension Buffer

Please do not hesitate to Contact us to find out more!

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Immuno-PCR Data

Immuno-PCR data generated using Thunder-Link®. A mouse monoclonal antibody specific for human CRP (clone C7) was purchased in unconjugated format from HyTest. The antibody was conjugated to an oligonucleotide using a Thunder-Link® kit, and was used as a detection antibody in a sandwich immuno-PCR assay with a polyclonal anti-CRP antibody as the capture reagent.

The top graph shows a plot of the number of PCR cycles undertaken vs. fluorescence intensity generated by SYBR Green containing PCR probes at particular antigen concentrations. The bottom graph shows this data following conversion to antigen amount vs. cycle number to enable the calculation of a standard curve. The results show that the assay utilizes 1000-fold less capture antibody and 100-fold less detection antibody, and provides 1000-fold greater sensitivity than the equivalent ELISA.

Frequently Asked Questions (FAQs) for Thunder-Link® PLUS

Thunder-Link® technology enables quick and easy conjugation of oligonucleotides to antibodies

1. What buffer should my oligonucleotide be in?

Suitable buffers must be between pH 6 and pH 8. The buffer must not contain primary amine groups. Additives such as BSA and azide should be avoided. Ideally the oligo will be dissolved in a phosphate buffer pH 7.

2. What size and modifications should my oligo have?

The oligo must be between 20 and 120 base pairs in length and contain a terminal amine group, which must be added during synthesis. All commercial oligo suppliers offer this modification. The efficiency of conjugation is slightly higher with 5’aminated oligos.

3. How pure should my oligo be, and at what concentration?

The oligo must be HPLC purified and be 60-100µM (high) concentration and be in 100µl of a suitable buffer (anything that is compatible with the downstream application). An option would be to resuspend the oligo in 100mM Phosphate buffer (pH 7.2). If the oligo concentration is greater than 100µM, dilute to 100µM in Wash Buffer.

4. What buffer should my antibody be in?

Suitable buffers must be between pH 7 and pH 9. The buffer must not contain primary amine groups. Additives such as BSA and azide should be avoided. Ideally the antibody will be dissolved in a phosphate buffer pH 7.

5. What are the other requirements for my antibody?

The antibody to be activated must be purified and at a concentration of 1mg/ml. Higher antibody concentrations should be diluted to 1mg/ml with wash buffer. With antibody concentrations below 1mg/ml the antibody must be concentrated before use.

6. How much antibody will the kit conjugate?

The kits are designed to activate 100µg of antibody in 100µl of a suitable buffer.

Conjugation efficiency is about 95-100%.

7. How do I store my new antibody/oligonucleotide conjugate?

The long term stability of the antibody-oligo conjugate will depend on a multitude of factors, including the antibody and oligo themselves, the storage temperature and conditions.

In order to maximise stability, we would recommend storing the conjugate as concentrated as possible and at a low temperature. We would suggest checking with the antibody and oligo manufacturers if their products can be stored in 50% glycerol at -20°C – most conjugates should be able to be stored in these conditions, as long as the antibody and oligo can handle -20°C. If it is appropriate for your reagents, the addition of preservatives may also be helpful.

8. What is the difference between Thunder-Link® and Thunder-Link® PLUS?

We have been listening to our Thunder-Link customers since we launched the product and have used their feedback and experiences to develop Thunder-Link® PLUS as a superior alternative to our original Thunder-Link technology. The protocol is much the same only faster; so now antibody and oligo activation only requires 30 minutes and the whole conjugation is finished in an hour compared to overnight as with the original Thunder-Link® technology. There is also an improved antibody and oligo recovery using Thunder-Link® PLUS and the clean-up procedure is more robust and flexible, now working with antibody fragments and small proteins too.

If you would like to find out more about Thunder-Link® PLUS please view the product here, or get in touch.

Please see here for ordering & shipping information.

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