Lightning-Link® Fluorescein Antibody Labeling Kit

Images

 
Flow Cytometry: Lightning-Link Fluorescein Antibody Labeling Kit [707-0010] - Mouse anti-duck CD4 antibody was conjugated with FITC using a Lightening-Link kit. The conjugated antibody was used to stain duck spleen and ...read more
Immunofluorescence: Lightning-Link Fluorescein Antibody Labeling Kit [707-0010] - RAW264 macrophage cell line stained with chicken anti-F4/80 antibody conjugated to Fluorescein using Lightning-Link®, ...read more

Product Details

Summary
Conjugate
FITC
Format
FITC (A=495, E=519)
Kit Type
Antibody Labeling Kit

Order Details

Lightning-Link® Fluorescein Antibody Labeling Kit Summary

Description
Lightning-Link antibody labeling kits enable the direct labeling of antibodies, proteins, peptides or other biomolecules for use in R&D applications, drug discovery and the development of diagnostic kits (See protocol for further information).

Our Fluorescein antibody labeling kit enables the direct conjugation of Fluorescein to any biomolecule with an available amine group. The researcher simply pipettes the antibody or other biomolecule into the vial of Lightning-Link Fluorescein and incubates for 3 hours.

FeaturesBenefits
Quick and easy to useSave time, no special knowledge required
No separation steps100% recovery - no antibody/protein loss
Can be used in a wide range of applicationsFlexible
Freeze driedShips at ambient temperature, long shelf-life
Fully scalable (10 ug to 1 g or more)Easy transfer from R&D to manufacturing
Stringently QC testedConsistent high quality, excellent batch-to-batch reproducibility
Large number of labels available Experimental flexibility
Reliable: nearly 300 referencesSuccessfully used in many fields of research


Fluorescein is one of the most popular fluorescent reagents used in biological research because of its water solubility, intense fluorescence and high absorptivity. It has a peak excitation occurring at 498nm and peak emission of 532nm.
Kit Type
Antibody Labeling Kit

Applications/Dilutions

Dilutions
  • Flow Cytometry
  • Immunofluorescence
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.
Reviewed Applications
Read 2 Reviews rated 5
using
707-0010 in the following applications:

Publications
Read Publications using
707-0010 in the following applications:

  • 9 publications
  • IA
    1 publication
  • 4 publications
  • IHC
    1 publication

Packaging, Storage & Formulations

Storage
Store at -20C.

Kit Components

Components
  1. 1 or 3 glass vial(s) of Lightning-Link mix
  2. 1 vial of LL-Modifier reagent
  3. 1 vial of LL-Quencher reagent

Notes

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

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.

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.

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) Fluorescein Kit (707-0010)(31)

We have publications tested in 1 confirmed species: Avian.

We have publications tested in 4 applications: FLOW, IA, ICC/IF, IHC.


Filter By Application
FLOW
(9)
IA
(1)
ICC/IF
(4)
IHC
(1)
All Applications
Filter By Species
Avian
(1)
All Species
Showing Publications 1 - 10 of 31. Show All 31 Publications.
Publications using 707-0010 Applications Species
Shanmugasundaram R, Wick M, Lilburn MS Effect of a post-hatch lipopolysaccharide challenge in Turkey poults and ducklings after a primary embryonic heat stress Dev. Comp. Immunol. Jul 5 2019 [PMID: 31283944] (Avian) Avian
Yang Y, Yeh SH, Madireddi S et al. Tetravalent biepitopic targeting enables intrinsic antibody agonism of tumor necrosis factor receptor superfamily members MAbs Jun 20 2019 [PMID: 31156033]
Micheva-Viteva SN, Ross BN, Gao J et al. Increased mortality in mice following immunoprophylaxis therapy with high dosage of nicotinamide in Burkholderia persistent infections. Infect. Immun. Oct 15 2018 [PMID: 30323029] (ICC/IF) ICC/IF
Shanmugasundaram R, Wick M, Lilburn M. Effect of embryonic thermal manipulation on heat shock protein 70 expression and immune system development in Pekin duck embryos. Poult Sci. 2018 Aug 14 [PMID: 30124990] (FLOW) FLOW
Do DC, Yang S, Yao X et al. N-glycan in cockroach allergen regulates human basophil function. Immun Inflamm Dis. 2017 [PMID: 28474843]
Pieper IL, Radley G, Chan CH et al. Quantification methods for human and large animal leukocytes using DNA dyes by flow cytometry Cytometry A. 2016 Jun [PMID: 27271958] (FLOW) FLOW
Yasmin AR, Yeap SK, Tan SW et al. In vitro characterization of chicken bone marrow-derived dendritic cells following infection with very virulent infectious bursal disease virus. Avian Pathol. 2015 [PMID: 26305169]
Khairalla AS, Omer SA, Mahdavi J et al. Nuclear trafficking, histone cleavage and induction of apoptosis by the meningococcal App and MspA autotransporters. Cell Microbiol 2015 [PMID: 25600171]
Rodgers JM, Robinson AP, Rosler ES et al. IL-17A activates ERK1/2 and enhances differentiation of oligodendrocyte progenitor cells. Glia 2014 [PMID: 25557204]
Robinson AP, Rodgers JM, Goings GE, Miller SD. Characterization of Oligodendroglial Populations in Mouse Demyelinating Disease Using Flow Cytometry: Clues for MS Pathogenesis. PLoS One 2014 [PMID: 25247590] (FLOW) FLOW
Show All 31 Publications.

Reviews for Lightning-Link (R) Fluorescein Kit (707-0010) (2) 52

Average Rating: 5
(Based on 2 reviews)

Reviews using 707-0010:
Filter by Applications
ICC/IF
(1)
All Applications
Filter by Species
All Species
Images Ratings Applications Species Date Details
Immunocytochemistry/Immunofluorescence Lightning-Link (R) Fluorescein 707-0010
Enlarge
5
reviewed by:
Revathi Shanmugasundaram
ICC/IF 12/12/2016
View

Summary

ApplicationImmunocytochemistry/Immunofluorescence

Comments

CommentsMouse anti-duck cd4 antibody was conjegated with this Fitc. It works fine with flowcytometry.
Serum Lightning-Link (R) Fluorescein 707-0010
Enlarge
5
reviewed by:
Revathi Shanmugasundaram
12/12/2016
View

Summary

ApplicationSerum

Comments

CommentsMouse anti-duck cd4 antibody was conjegated with this Fitc. It works fine with flowcytometry.

Product General Protocols

View specific protocols for Lightning-Link (R) Fluorescein Kit (707-0010):
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Lightning-Link (R) Fluorescein Kit (707-0010). (Showing 1 - 10 of 10 FAQs).

  1. I want to label an anti-human AB with your Lighning Link FITC labeling kit and use it then for FACS analyses. How long will the labeling of the antibody be stable? Can the labeled antibody be used for FACS
    • The labeled antibody will be stable for as long as the life of the antibody, provided that it is properly stored in the dark. I recommend using a brown opaque tube and storing your antibody at 4degC. It will also depend on your antibody whether it can be used for FACS. There is nothing in the labeling reaction that will inhibit the antibody from being used in FACS, but its performance may depend on the antibody itself.
  2. I want to label an anti-human AB with your Lightning Link FITC labeling kit and use it then for FACS analyses. How long will the labeling of the antibody be stable? Can the labeled antibody be used for FACS?
    • The labeled antibody will be stable for as long as the life of the antibody, provided that it is properly stored in the dark. I recommend using a brown opaque tube and storing your antibody at 4C. It will also depend on your antibody whether it can be used for FACS. There is nothing in the labeling reaction that will inhibit the antibody from being used in FACS, but its performance may depend on the antibody itself.
  3. 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.
  4. 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.
  5. 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.
  6. 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.
  7. 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.
  8. 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.
  9. 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.
  10. 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.

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Recent Reviews

5
5
2
4
0
3
0
2
0
1
0

Revathi Shanmugasundaram
12/12/2016
Application: ICC/IF
Species:

Revathi Shanmugasundaram
12/12/2016
Application:
Species: