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Oligonucleotide Conjugation Kits

Thunder-Link®

Easy to use Conjugation Kits for Oligonucleotides

 

Thunder-Link PLUS

 

 

Thunder-Link® technology enables quick and easy conjugation of oligonucleotides to antibodies for use in R&D applications, drug discovery and the development of diagnostic kits.

Using standard chemistry and methodology, the linking of an oligonucleotide to molecules is difficult, time consuming and requires specialist knowledge. To overcome these issues the Thunder-Link® oligo conjugation system has been designed to allow both specialists and non-specialists to generate antibody oligonucleotide conjugates easily and efficiently. Furthermore:

  • Technology is unidirectional and thus allows formation of molecule-oligonucleotide complexes only and never molecule-molecule or oligonucleotide-oligonucleotide.
  • Any oligo sequence may be used and the target chemistry may be at the 5' or 3'.
  • Thunder-Link® for linking oligos to antibodies is provided with a positive control to demonstrate successful conjugation.

Thunder-Link® PLUS allows even faster and more efficient antibody-oligonucleotide conjugation than our original Thunder-Link® kit (discontinued in May 2016). We have developed this new superior kit to enable oligo conjugation in only an hour with higher recovery of materials.  Find out more here.

 

The Thunder-Link® conjugation process

immuno pcr, oligo antibody conjugation

 Contact us to find out more:

Thunder-Link® PLUS Oligonucleotide Conjugation Kit [Find out more]

 

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.

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.

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. If the oligo concentration is greater than 100µM, dilute to 100µM in Wash Buffer.

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.

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.

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%.

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.

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 at info@innovabiosciences.com.

"We have been working with Innova for about a year now and have had nothing but fantastic experiences with both their sales staff as well as their R&D staff. From the timely generation of quotes and custom project pricing, to the extremely helpful discussions with R&D, we have been duly impressed. Everyone on the Innova team handles themselves with the utmost professionalism and courtesy, and are experts in field.

Regarding custom antibody:oligonucleotide conjugates, we were initially impressed with the quality of the conjugates we ordered, and are constantly being “re-impressed” since the quality of subsequent conjugates keeps improving! Again, the Innova team’s expertise in the challenging field of antibody conjugation allows them to tackle any project we toss to them with no hesitation and generate a top-notch product to boot. Overall, we are thoroughly pleased with Innova’s work and staff. We would whole-heartedly recommend them to anyone who needs excellent custom labeling and conjugation work – you will not be disappointed!"

Lou Casta, Ph.D. at Genisphere, LLC.

Immuno-PCR

The detection of rare cells or low concentrations of proteins with accuracy and sensitivity in test samples such as whole blood and serum is a significant problem. This is especially true with standard immunoassays such as ELISA.

With the amplification offered by immuno-PCR, the detection of single nucleic acid molecules is now routine -a DNA probe hybridized to its template offers high sensitivity and specificity. The next generation of immunoassay reagents are being engineered through linking oligonucleotides to a diverse set of molecules for instance antibodies or gold nanoparticles.

Typically oligonucleotide conjugates are used in immuno-PCR which combines the specificity of antibodies with the amplification power of immuno-PCR allowing a 10 to 1000 fold increase in sensitivity. Oligonucleotide conjugates also have the potential to be the platform tool in multiplexed protein diagnostic assays.

ThunderLink Immuno-PCR

For more information, take a look at our Immuno-PCR application page.

Proximity ligation assay

Used to identify proteins co-located within cells

ThunderLink ProximityLigationAssay

Proximity extension assay

Variation on ImmunoPCR using two antibodies specific for different epitopes

ThunderLink ProximityExtensionAssay

Fluorescent antibody labeling

Directly fluorescent oligos or complementary labels

  

 

You can find out more about oligo-antibody conjugates from our recent webinar - Oligo:Ab conjugates - an application guide.

 



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