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Lightning-Link®

Lightning-Link® is an innovative technology that enables direct labeling of antibodies, proteins, peptides or other biomolecules for use in R&D applications, drug discovery and the development of diagnostic kits.

The kits:

  • Require only 30 seconds hands-on time
  • Do not require spin or separation steps
  • Are easy to use

The researcher simply pipettes the antibody or other biomolecule into a vial of lyophilized mixture containing the label of interest and incubates for either 3 hours (Lightning-Link range) or only 15 min(Lightning-Link Rapid range). Despite its apparent simplicity, the Lightning-Link® process is sophisticated and generates conjugates with performance characteristics identical to, or better than, those prepared with laborious multistep conjugation procedures.

The Lightning-Link® range currently consists of 50+ kits including enzymes, biotin, streptavidin and fluorochromes covering the spectrum from UV to far infrared. Lightning-Link® conjugates are used in a large number of scientific applications including Western Blotting, immunofluorescence, immunohistochemistry, flow cytometry, ELISA and FRET.

The technology is fully scalable (µg to grams) ensuring quality and consistency without any deterioration in the performance of the conjugate. As there are no separation or desalting steps, antibody recovery is 100%. Furthermore, the bond between the label and biomolecule is covalent, therefore the conjugate is extremely stable.

Testimonials

VolitionRx

Dr Jake Micallef, Chief Scientific Officer

We’ve been really impressed with Innova’s products.

Their biotin kit is simple to use and proved more durable than many competitors’ kits, and we’ve seen significant improvements in reproducibility since we started to use them regularly.

We’re looking forward to continuing to work with the Lightning–Link® technology.

Bio-Check (UK) Limited

Phil Goodwin, Managing Director

Like many other small diagnostics companies we have used our own HRP activation and conjugation techniques for some years. However, on trying Innova’s Lightning Link kit we found its simplicity and the high level of functionality of the conjugates produced to be significantly better. We have since tried another, similar kit but the resulting product was decidedly inferior, with lower signal and higher noise. We plan to continue using Lightning–Link® for our existing ELISAs and also for others to be added to our food testing range in the next few years.

Tommy Tong, Post-Doctoral Fellow

Lightning–Link® Alkaline Phosphatase kit is a simple, easy-to-use kit, to biochemically label protein of interest, in our case, anti-HIV envelope antibodies, for ELISA. Labeling antibody with this kit does not affect the function and avidity of the antibody. One advantage in using this kit is that there is no need to remove excess alkaline phosphatase from the reaction mixture at the end of the incubation. The omission of this step does not affect our ELISA analysis.

FAQs

  • Which label should I use?

    Answer: Roughly, the Lightning-Link kits can be grouped into 3 categories: enzymes, biotin/streptavidin etc, and fluorescent dyes/proteins. Our product application table shows the most common labels used for certain applications, and our Fluorescent labeling table compares absorption and emissions of these kits. The individual product pages can then be used to narrow down which product is best for you.

  • Where does the label bind on my antibody?

    Answer: Unfortunately we cannot guarantee the answer to this question, as we are not able to control the labeling reactions. The active groups will target free amines on the biomolecule, but we cannot be specific as to which one. The most likely case is that the label will bind to the FC region, purely because there are more amine groups in this area. Theoretically, the label could bind to the binding site of the antibody, however this is statistically very unlikely, and we have hardly ever seen this happen.

  • What is the difference between Lightning-Link® and Lightning-Link® Rapid?

    Answer: The conjugate quality obtained by the two types of kits is identical. The only difference is the time taken for the conjugates to be ready. Our standard kits require a 3 hour incubation time, whereas the rapid kits only require 15mins. Most of our fluorescent dyes are only available in the rapid format.

  • What if I need bulk material?

    Answer: Our 'off-the-shelf' kit sizes are relatively small to allow testing for R&D purposes. If you require larger pack sizes, or large volumes of our existing kit sizes, this is not a problem. Please simply request a quote using the 'Custom Quote request' button, or contact our Technical Support Team at technical.enquiries@innovabiosciences.com to dicuss your requirements.

  • What are the benefits of Lightning Link over traditional labelling methods?

    Answer: The Lightning-Link® mix contains all of the necessary chemicals to allow you to conjugate your primary antibody directly to the label of your choice withjust 30 seconds hands on time. As the label is directly conjugated to your primary antibody, non-specific binding arising from secondary antibodies is eliminated in your assay. Also, as the by-products of the reaction are benign, there is no need for purification steps after the conjugation, meaning antibody recovery is virtually 100%. We have over 35 labels available to suit you needs.

  • What size kit should I buy?

    Answer: The answer to this will depend on how much antibody you wish to label, and which label you wish to use. As a vague rule, we provide three standard sizes of kits, 3 x 10ug, 3 x 100ug and 1 x 1mg, which are designed to label 'around' this amount of antibody. The exact amounts possible are detailed in the protocol for the chosen label, so please ensure to consult the relelvant document carefully before ordering. In short, our 3 x 10ug kits are designed for optimisation purposes. The three vials contained in the kit allow you to experiment with concentrations to identify the best ratio for your assay, using minimal amounts of antibody. The larger kit sizes are then available for labeling more of the antibody, once the best ratio has been determined. If a variation or a customised kit size would benefit you, please do not hesitate to contact our Technical Support Team to discuss this.

  • How stable will my new conjugate be?

    Answer: The Lightning-Link® chemistry joins the label to the antibody via a uni-directional, stable, covalent bond. The stability of the conjugate will therefore vary slightly depending on the antibody and label in use. In our in house studies, conjugates were still fully active after more than 18 months storage. The necessary storage conditions varies depending on the label and antibody. Please consult the protocol for the relevant Lightning-Link® kit for further information.

  • How many labels will bind to my biomolecule?

    Answer: It is difficult to give a precise answer to this as the result will vary from antibody to antibody (or other protein), and between different labels. Traditional labelling techniques with small dyes e.g. fluorescein, often have F/P ratios in the range of 4-7:1, and our comparative data for staining with antibodies labelled with Lightning Link shows similar staining intensity. For fluorescent proteins, e.g. R-PE, the typical labelling ratio is 1:1. The ratio of dye to antibody is only ever an average for the population of labelled molecules, as individual molecules may have different amount of dye incorporated into them. In most cases. the number of labels per molecule is outlined in the individual protocols.

  • Do my antibody and buffer fit the requirements?

    Answer: In order for your antibody to succesfully conjugated, it needs to be purified, and in a suitable, amine free buffer. Details of contaminating substances, and suitable buffer requirements can be found on the protocol for the Lightning-Link® product.

  • Do I need a wash or desalt step?

    Answer: No. The Lightning-Link® technology has been optimised so that when the protocols are followed correctly, there is virtually no free label remaining. Any labels that are left over have there reactive groups blocked by the quencher step, meaning they will not interfere. They are then simply washed away by the wash steps in your chosen application.

  • Can I label a peptide/protein/antibody other than IgG?

    Answer: Yes. The Lightning-Link® technology conjugates to free amine groups on your target, meaning it can be used to label most biomolecules. However the protocol may need adapting slightly. See the protocol for your chosen label to see any necessary amendments.

  • Can different sub-types of antibodies be used?

    Answer: Yes, however a slight protocol adapatation may be require dependant on size difference. Our protocols are optimised for labeling IgG antibodies, so we would recommend adapting the amounts involved according to the size difference of your subtype vs. IgG. Please see the protocol for your chosen label for details on how to accomodate this. Please note: although IgM antibodies are much larger than IgG's, we recommend that you follow the protocol as though you were labelling IgG, as this usually gives the best results. The best amount for your particular antibody will need to be determined experimentally.

  • Can antibody fragments be conjugated?

    Answer: Yes, as they contain amine groups, this will not be a problem. We have seen many succesful conjugations of Lightning-Link® with antibody fragments in the past. The protocols are optimised for labeling whole IgG molecules, so please consult the protocol of your chosen label for details on how to accomodate for difference in size.

  • Can antibodies from different species be used?

    Answer: Yes, as the chemistry is not specific to a specie, but works by tragetting primary amine groups on your molecule. Antibodies from all species will contain amine groups, and are therefore compatible with Lightning-Link.

  • Do I need to purify the conjugate post conjugation?

    Answer: No. The chemicals used in Lightning-Link® and Lightning-Link® Rapid kits are deactivated by the quencher provided in the kits, and the by-products are benign and do not need to be removed.

  • What are the best storage conditions for my new conjugate?

    Answer: The storage conditions vary depending on the label used. Please consult the protocol for the label of choice to identify the best storage conditions. These will also vary depending on the antibody, so please bear this in mind.