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Antibody Labeling Kits

Lightning-Link®

For labeling primary antibodies, proteins or peptides

 

Primary antibody and protein labeling

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.

Benefits of Lightning-Link® include:

  • Quick and easy to use
  • Requires only 30 seconds hands-on time
  • No separation steps involved so recover 100% of your antibody or protein
  • Label from as little as 10ug to a gram or more!

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)View Lightning-Link Rapid range>>

LL Process diagram-new

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.

To learn more, read our introductory guide to antibody labeling.

Contact us to find out more:

 

Manufacturing

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.

For more information click here >>

    • "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.”

      Dr Jake Micallef, CSO of VolitionRx

 

    • "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.”

      Chief Executive at Bio-Check (UK) Limited

 

  • See what others have said here!
Lightning-Link® conjugates are used in a large number of scientific applications including:
We also have a range of educational videos available, please follow the links below:
 

Which label should I use?

The choice of label for your antibody primarily depends upon the application:

Immunoassay Label
Western Blotting
  • Enzymes (usually HRP or Alkaline Phosphatase)
ELISA
  • Enzymes
  • Biotin/Streptavidin
Immunofluorescence
  • Fluorescent dyes
Immunohistochemistry
  • Enzymes
  • Biotin/Streptavidin
Flow Cytometry
  • Fluorescent proteins
  • Fluorescent dyes
  • Tandem dyes

Please see the table here for absorption/emission wavelengths of Lightning-Link® fluorescent dyes, proteins and tandems.

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

Both Lightning-Link® and Lightning-Link® Rapid kits can generate conjugates in a hands-on time of typically 20-30 seconds for the entire procedure. However the conjugation and quenching incubation times are shorter for Lightning-Link® Rapid kits allowing the conjugates to be generated faster. The antibody considerations are exactly the same, as is the high quality of the final conjugate.

What can I label using Lightning-Link?

Lightning-Link® technology works by targeting free amine groups on your target. It can be used to label antibodies, peptides, proteins and any other molecules with free amine groups.

For advice on labeling molecules other than antibodies please contact our technical support team.

How pure does my antibody need to be?

Your antibody must be purified because other molecules with free amine groups will interfere with the reaction resulting in a poor quality conjugate.

A suitable method of purification should be selected:

  • Affinity purification - this is the preferred method as it will always result in a purified sample containing only the antibody of interest.
  • Protein A or G purification of serum - this will result in a purified sample of all IgG’s contained in the serum. You will have a purified IgG fraction but only a small percentage of this fraction will be the IgG of interest. This IgG fraction will label with our kits, although adjustments in conjugate dilutions will be required.
  • Protein A or G purification of tissue culture supernatant or ascites fluid - this method will generate purified antibodies equivalent to affinity purification.

Other purification methods, such as ion exchange chromatography and ammonium sulphate precipitation, are low quality purification methods and should be avoided. Filtration using a 0.2/0.22 µm filter is a method of sterilization not purification, and is therefore not appropriate.

Is my antibody in a suitable buffer?

For recommended buffer conditions please see the below table:

Buffer components
pH 6.5-8.5
Amine free buffer (e.g. MES, MOPS, HEPES, PBS) ü
Non-buffering salts (e.g. sodium chloride) ü
Chelating agents (e.g. EDTA) ü
Sugars ü
Glycerol <50%
Thiomersal/Thimerosal x
Merthiolate x
Sodium Azide1,3 <0.1%
BSA1,2 <0.1% 
Gelatin1,2 <0.1% 
Tris <50mM
Glycine
Proclin
Borate buffer ü 
Nucleophilic components (Primary amines e.g. amino acids or ethanolamine and thiols e.g. mercaptoethanol or DTT) x

 1 Please note that individually the concentrations shown should not affect the reaction. However in combination with additional compounds that are not recommended above a certain concentration, the reaction may be affected.

2 If intending to use this kit for immunohistochemistry, it is recommended that there be no gelatin or BSA present.

3 It is important to note that sodium azide is a known inhibitor of HRP and should be avoided.

What can I do if my antibody formulation does not fit the requirements?

If your antibody requires purification or the buffer is not compatible with our kits, we have developed an AbSelect™ purification kit range that allows you to quickly and simply purify your antibody and is fully compatible with the Lightning-Link® kits.

The appropriate kit to use depends on your particular sample (species, buffer, contaminants, volume, etc.). We have designed a handy flow chart on the AbSelect™ webpage to help you select a kit, but feel free to contact us if you require further guidance. Please consult the individual kit protocols to see the antibody amount/volume suitable for each kit.

If your antibody is already purified but its concentration is significantly lower than 1mg/ml, you can concentrate it using our Antibody Concentration and Clean Up Kit. This kit can also be used to remove low molecular weight contaminants such as azide, tris or glycine by carrying out a buffer exchange into the buffer supplied in the kit, which is fully compatible with Lightning-Link®.

If your antibody contains BSA, you can now use our BSA removal kit to purify your antibody in a few simple steps. Please note this kit will also enable you to concentrate your antibody.

NB: All the AbSelect kits will ONLY work with antibodies. They will not purify other molecules. The only exception is the concentration and clean up kit (861-0010), which will work with other molecules greater than 10kD.

Do I need a wash or desalt step?

One of the advantages of the Lightning-Link® technology compared to traditional labeling methods is that conjugate purification is not required. The Lightning-Link® reactions are highly efficient, and the kits have been optimized so that, provided the protocols are followed, there should be only a very low amount of free label left at the end of the conjugation. This remaining free label would have its reactive 'Lightning-Link®' groups blocked by the Quencher provided in the kit, and is washed away during the wash step of your application.

How stable will my new conjugate be?

Upon dissolution of the Lightning-Link® mixture with a solution of the antibody, proprietary chemicals in the mixture become activated. This results in the directional covalent bonding of the antibody to the label in a gentle and controlled process at near-neutral pH. The stability of your conjugate is dependent on your antibody and label of choice.

Most Lightning-Link® conjugates can be stored for up to 18 months, undiluted at 4°C. The exception to this is tandem conjugates which are only stable for up to 3 months.

 

 
Lightning-Link ProductsClick the individual products for further information, pricing and data.

Enzyme Labeling Kits

Biotin / Streptavidin Labeling Kits

Fluorescence Labeling Kits

Lightning-Link®*
Lightning-Link® Rapid*
Maximal Absorbance
(nm)
Maximal Emission
(nm)
AMCA
 
352
452
DyLight® 350
 
354
432
Atto 390
 
388
468
DyLight® 405
 
402
428
PerCP 718-0030 484 678
PerCP/Cy5.5 763-0030 484 692
DyLight® 488   322-0030 496 524
Fluorescein 498
532
R-Phycoerythrin
498,544,566**
580
PE/Texas Red 767-0030 498,544,566** 618
PE/Atto594 768-0030 498,544,566** 632
PE/Cy5 760-0030 498,544,566** 672
PE/Cy5.5 761-0030 498,544,566** 700
PE/Cy7 762-0030 498,544,566** 782
Atto 488   350-0030 504 530
B-Phycoerythrin
546
580
Cyanine Dye 3 (Cy3) 340-0030 552 576
Rhodamine
555
588
DyLight® 550
 
556
584
Atto 565
 
570
598
DyLight® 594
 
594
629
Texas Red®
 
596
616
DyLight® 633
 
628
660
Atto 633 353-0030 634 660
FluoProbes647H 362-0030 650 684
Cyanine Dye 5 (Cy5)
 
652
678
Allophycocyanin
 
652
666
APC/Cy5.5 764-0030 652 700
APC/Cy7
 
652
790
DyLight® 650
656
686
Cyanine Dye 5.5 (Cy5.5)
 
680
705
DyLight® 680
 
686
716
Atto 700
 
704
724
DyLight® 755
 
756
794
DyLight® 800
 
776
798
* The antibody labeling kits come in two formats - Lightning-Link® and Lightning-Link® Rapid, with incubation times of 3 hours and 15 minutes respectively.
**(R-)PE has three maxima, and all can be used. The optimal will depend on the application.

For a downloadable version of the fluorescence table click here>>

Fluorochrome guide download PDF

by InnovaBiosciences 




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