Thunder-Link® PLUS FAQs
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 10 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.