Lightning-Link® Rapid DyLight 488 Antibody Labeling Kit

Flow Cytometry: Lightning-Link (R) Rapid DyLight 488 Antibody Labeling Kit [322-0010] - DL488 labeled virus detected with antibody-coated beads. Flow cytometry image submitted by a verified customer review.

Flow Cytometry: Lightning-Link Rapid DyLight 488 Antibody Labeling Kit [322-0010] - Mouse anti-human CD3 was conjugated with Dylight® 488 using Lightning-Link ® Rapid kit. The conjugated antibody was then used to stain human peripheral blood lymphocytes, followed by analysis with flow cytometry. (Blue line - negative control; red line - positive staining).

Flow Cytometry: Lightning-Link Rapid DyLight 488 Antibody Labeling Kit [322-0010] - Mouse anti-duck CD4 was conjugated with DyLight 488 using a Lightening-Link Rapid kit. The conjugated antibody was used for flow cytometry. Image from verified customer review.

Lightning-Link Rapid DyLight 488 Antibody Labeling Kit [322-0010]

• Conjugate: DyLight 488
• Format: DyLight 488 (A=493, E=518)
• Kit Type: Antibody Labeling Kit

Lightning-Link® Rapid DyLight 488 Antibody Labeling Kit Summary

• Description: Lightning-Link Rapid is an innovative technology that enables direct labeling of proteins, peptides or other biomolecules for use in R&D applications, drug discovery and the development of diagnostic kits (See protocol for further information).
  
  The easy-to-use, one step procedure allows researchers to covalently label biomolecules with only 30 seconds hands-on time; furthermore conjugates are ready to use in less than twenty minutes.
  
  The researcher simply pipettes the biomolecule into a vial of lyophilized mixture containing the label of interest and incubates (for more details please watch the video below).
  
  

  Features	Benefits
  Quick and easy to use	Save time, no special knowledge required
  No separation steps	100% recovery - no antibody/protein loss
  Can be used in a wide range of applications	Flexible
  Freeze dried	Ships at ambient temperature, long shelf-life
  Fully scalable (10 ug to 1 g or more)	Easy transfer from R&D to manufacturing
  Stringently QC tested	Consistent high quality, excellent batch-to-batch reproducibility
  Large number of labels available	Experimental flexibility
  Reliable: nearly 300 references	Successfully used in many fields of research

  
  
  DyLight 488 provides green fluorescence for a wide array of fluorescence labeling-based applications. It has a strong absorption at 496nm, high fluorescence at 524nm (extinction coefficient 7.0 x104 cm-1M-1) and high quantum yield.
  
  Learn more about Lightning-Link™ Conjugation Kits by reading FAQs
  
  For more information please check out these useful links!
  Antibody Labeling Guide
  Antibody Conjugation Illustrated Assay
• Kit Type: Antibody Labeling Kit

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
• Application Notes: By circumventing the desalting or dialysis steps that commonly interrupt traditional antibody conjugation procedures, LightningLink technology can be used to label both small (e.g. 10 ug) and large quantities of primary antibodies with ease. Batch-to-batch variation upon scale up is minimal as the process is so simple, and recoveries are always 100%. This kit is supplied with 3 vials, each suitable for labeling up to 200 ug of antibody.

Packaging, Storage & Formulations

• Storage: Store at -20C.

Notes

This product is manufactured by Abcam and distributed by Novus Biologicals.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. This product is guaranteed for 1 year from date of receipt and this statement overrides any mentioned guarantee period on the limitations section of this products datasheet. Please contact technical@novusbio.com with questions.

Background

Lightning-Link® Rapid Antibody Labeling kits

Allows direct labeling of antibodies, proteins, peptides or other biomolecules. Procedure is completed in seconds, and used within minutes (less than 20 minutes). Process generates conjugates identical, or better than, traditional conjugation procedures. 100 percent recovery.

For more information please check out these useful links!

Antibody Labeling Guide

Purifaction Antibody Guide

Lightning-Link® Demonstration Video

Fluorescent Labels Poster

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for Lightning-Link (R) Rapid DyLight 488 Kit (322-0010)(14)

Publications using 322-0010

• Keeley EC, Schutt RC, Marinescu MA et al. Circulating fibrocytes as predictors of adverse events in unstable angina Transl Res. 2016-03-08 [PMID: 27012475] (FLOW)
• Crawford JR, Trial J, Nambi V et al. Plasma Levels of Endothelial Microparticles Bearing Monomeric C-reactive Protein are Increased in Peripheral Artery Disease J Cardiovasc Transl Res. 2016-02-18 [PMID: 26891844] (FLOW)
• Kerkela E, Laitinen A, Rabina J et al. Adenosinergic immunosuppression by human mesenchymal stromal cells (MSCs) requires co-operation with T cells Stem Cells 2016-03-01 [PMID: 26731338] (FLOW)
• Cumpelik A, Gerossier E, Jin J et al. Mechanism of Platelet Activation and Hypercoagulability by Antithymocyte Globulins (ATG) Am J Transplant. 2015-10-01 [PMID: 25966640] (FLOW)
• Wu Y, Simons J, Hooson S et al. Protein and virus-like particle adsorption on perfusion chromatography media. J Chromatogr A. 2013-01-01 [PMID: 23726244]
• Leetah Celine Senkpeil Identifying Immunological Signatures in Blood Predictive of Host Response to Plasmodium falciparum Vaccines and Infections Using Computational Methods Thesis 2023-01-01
• Leetah Celine Senkpeil Identifying Immunological Signatures in Blood Predictive of Host Response to Plasmodium falciparum Vaccines and Infections Using Computational Methods Thesis 2023-01-01
• Wimmers F, Donato M, Kuo A et al. The single-cell epigenomic and transcriptional landscape of immunity to influenza vaccination Cell 2021-07-01 [PMID: 34174187]
• Hollandsworth HM, Schmitt V, Amirfakhri S et al. Fluorophore-conjugated Helicobacter pylori recombinant membrane protein (HopQ) labels primary colon cancer and metastases in orthotopic mouse models by binding CEA-related cell adhesion molecules Transl Oncol 2020-08-28 [PMID: 32866936]
• Garreta E, Prado P, Tarantino C et al. Fine tuning the extracellular environment accelerates the derivation of kidney organoids from human pluripotent stem cells Nature Materials 2019-04-01 [PMID: 30778227]
• Iwao Y, Tomiguchi I, Domura A, Mantaira Y. Inflamed site-specific drug delivery system based on the interaction of human serum albumin nanoparticles with myeloperoxidase in a murine model of experimental colitis. Eur J Pharm Biopharm. 2018-01-30 [PMID: 29407223] (FLOW)
• Simanjuntak Y, Liang JJ, Lee YL et al. Japanese Encephalitis Virus Exploits Dopamine D2 Receptor-phospholipase C to Target Dopaminergic Human Neuronal Cells. Front Microbiol 2017-04-11 [PMID: 28443089]
• Mellema M, Stoller M, Queau Y et al. Nanoparticle Tracking Analysis for the Enumeration and Characterization of Mineralo-Organic Nanoparticles in Feline Urine. PLoS One. 2016-12-22 [PMID: 28005930]
• Schmiedel D, Tai J, Levi-Schaffer F et al. Human Herpesvirus 6 downregulates the expression of activating ligands during lytic infection to escape elimination by natural killer cells. J Virol. 2016-10-14 [PMID: 27535049]

Reviews for Lightning-Link (R) Rapid DyLight 488 Kit (322-0010) (2)

Average Rating: 5

(Based on 2 reviews)

reviewed by: Verified Customer 03/04/2020

Summary

• Lot: IBS4346

reviewed by: Revathi Shanmugasundaram 12/12/2016

Comments

• Comments: Conjugated with mouse antiduck cd4 antibody. works fine with flowcytometry.

FAQs for Lightning-Link (R) Rapid DyLight 488 Kit (322-0010). (Showing 1 - 10 of 10 FAQs).

1. Can I label a peptide/protein/antibody other than IgG?
   • Yes. The Lightning-Link® technology works by targeting free amine groups on your target, meaning it can be used to label most biomolecules.As the protocols provided were optimized for labeling IgGs, we would recommend you adjust the amount of material you add to the Lighting-Link® vial to allow for molecular weight difference. This should be done without changing the volume added to the vial, as this could affect the conjugation efficiency. As a rough guideline, we would recommend changing the amount of material proportionally to the size difference with IgGs. An average IgG is about 160kD, therefore for a target that is half the size of an antibody (about 80kD), add half as much to the vial. Please note this is only a guideline and the best amount for your assay should be determined experimentally; our 3x10ug kits enable you to do this using small amounts of material and therefore at a low cost.
2. 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. Any remaining free label would have its reactive 'Lightning-Link®' groups blocked by the Quencher provided in the kit, and would then be washed away during the relevant wash step of your application.
3. Do my antibody and buffer fit the requirements?
   • Your antibody should be purified (affinity purification is preferred), as other molecules with free amine groups will interfere with the reaction, resulting in a poor quality conjugate. A suitable method of purification should be used (such as affinity or Protein A/G). Please note that 0.2/0.22 um filtration is a method of sterilization, not purification.Your biomolecule should also be in a suitable, 10-50mM amine free buffer (e.g. MES, MOPS, HEPES, PBS), pH range 6.5 to 8.5 and not in any of the following: ascites fluid, serum or tissue culture supernatant. It should not contain any additives such as Azide, BSA, Tris or Glycine at a concentration of >0.1%.The amount of antibody (IgG) you should add to the Lightning-Link® vial usually corresponds to the kit size your purchased (for example: a 3x100ug kit enable you to label 3 lots of 100ug antibody) and the volume added should also match (eg: 100ul), meaning the ideal concentration is of 1mg/ml. For other biomolecule, the amount added should be adjusted by changing your concentration (see above).NOTE: The amount and volume of antibody recommended above are for all Lightning-Link• kits offered by Innova Biosciences, with the exception of RPE and PE tandem dyes. For more details on the recommendations specific to these dyes, please consult the protocols available on each product page.
4. How many labels will bind to my biomolecule?
   • It is difficult to give a precise answer to this as the result will vary from antibody to antibody (or other protein). Traditional labelling techniques 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. 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.
5. How stable will my new conjugate be?
   • The Lightning-Link® chemistry joins the label to the antibody via a uni directional stable, covalent bond. The stability of your conjugate will therefore be dependent on your antibody and label of choice. In our in-house studies, conjugates made with our Lightning-Link® kits were fully active after more than 18 months' storage, undiluted, at 4 degrees C. Conjugates stored in 50% glycerol at -20 degrees C were found to be even more stable (several years). Please see below for advice on storage conditions. Please note that tandem dye stability is lower than the other dyes'. Tandem conjugates are fully stable for about 3 months at 4 degrees C.
6. What are the best storage conditions for my new conjugate?
   • TA new conjugate can be stored for 12-18 months at 4 degrees C as long as the antibody will tolerate storage at 4 degrees C. As the bond between the antibody and dye is covalent and very stable, it should not degrade, therefore at 4 degrees C no additional preservatives are needed. The antibody is usually the least stable component of the conjugate so please check the antibody datasheet for storage recommendations. However, all conjugates can be stored in 50% glycerol at -20 degrees C which allows them to remain stable for 2 years. Please bear in mind that APC and RPE conjugates should never be stored at -20 degrees C on their own without glycerol. The dilution factor also has an effect. Storing the conjugate undiluted is recommended if possible; however, for HRP conjugates, if you wish to store at working concentrations (e.g. 1/10,000), this can be done using our LifeXtend HRP Conjugate Stabilizer/Diluent. Please note that the preservative sodium azide will inhibit HRP. We suggest that the optimal storage conditions for any conjugate are determined experimentally, using small aliquots of the conjugate.
7. What can I do if my antibody formulation does not fit the requirements?
   • If your antibody 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,...). 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 kit protocols to see the antibody amount/volume suitable for each kit.If your antibody is already purified but its concentration is too low, you can concentrate it by 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.
8. What is the difference between Lightning-Link® and Lightning-Link® Rapid?
   • The benefit of the Rapid kits is that the conjugates can be generated faster because the conjugation and quenching incubation times are shorter. The antibody/protein/peptide considerations are exactly the same, as is the high quality of the final conjugate. DyLight® dyes are only available in the Rapid format.
9. What functional groups do I need on my protein?
   • Lightning-Link Rapid requires amine groups on the molecule to be labeled. Most proteins have lysine and/or alpha-amino groups. All antibodies will have multiple amine functions.
10. Do I need to purify the conjugate?
    • No. The chemicals used in Lightning-Link Rapid are deactivated by the Quencher, and the by-products are benign and do not need to be removed.