What functional groups do I need on my protein?

Lightning-Link 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.

Do I need to purify the conjugate?

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

Can non-antibody molecules be labeled?

Yes. While labeling of antibodies is a major application, the only requirement is that the protein to be labeled has amine functionality.

I have 2 vials of pure lyophilized TLR1 antibody (catalog# AF1484) and I want to label the antibodies with PE Flourochromes. I am not sure how many kits to order from Lightning Antibody Labeling Kits and if any antibody purification or concentration kit is required.

For our PE Lightning Link Conjugation kits, I would recommend purchasing the kit 703-0010 (suitable for conjugating 3 x 60ug of antibody). This is the kit that can conjugate the closest amount of antibody to the full quantity you have per vial. The next size up is suitable for conjugating 600ug of antibody per reaction, but is priced much higher than the 60ug reactions. For your reference, the 600ug kits are available as 703-0015 (1 x 600ug) or 703-0003 (5 x 600ug).

Is trehalose accepatable in PBS for label with Lightning Link R-PE

The trehalose should not affect any protein modification. Trehalose is a common component as lyophilised excipient and we do not expect that it will interfere with the conjugation.

I had discussions with technical experts about residual fluorescence of unreacted dye after protein modification using lightening link (PE). I have been repeatedly ensured that the un reacted dye looses fluorescence upon quenching. This has subsequently been determined to be untrue. Am I interpreting your answers correctly?

Yes you are correct. The quencher does not stop the fluorescence or block the label. It blocks the reactive amine group from being able to link to the free amines. The process in theory is called quenching because the label can no longer bind and, therefore, the unbound label (in most applications) will be washed away in the first wash step. We do not use this label kit for an antibody or protein in an application such as your cell labeling. If you want to use this kit in cell labeling you will want to do a separation from the unincorporated label. If you would like to use a different labeling chemistry or separate the unincorporated by size exclusion chromatography then you can contact our Custom service https://www.rndsystems.com/services/protein-services

Lightning-Link (R) R-PE Antibody Labeling Kit

Name: Lightning-Link (R) R-PE Antibody Labeling Kit
Brand: Novus Biologicals
SKU: 703-0010
Rating: 4 (3 reviews)

Novus Biologicals, part of Bio-Techne | Catalog # 703-0010

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Product Summary
Product Specifications
Scientific Data
Kit Contents
Preparation & Storage
Background
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Specific Notices

Key Product Details

Applications

Flow Cytometry

Kit Type

Antibody Labeling Kit

Conjugate

R-Phycoerythrin

Product Summary for Lightning-Link (R) R-PE Antibody Labeling Kit

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

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

R-Phycoerythrin (R-PE) is a fluorescent protein from the phycobiliprotein family, present in red algae and cryptophytes. It has three maximal absorbance values of 498, 544 and 566nm (the optimal will depend on the application), and it has a strong emission peak at 580nm. RPE is closely related to B-Phycoerythrin (B-PE) and these are the most intense fluorescent phycobiliproteins providing an orange fluorescence.

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

Product Specifications

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 60 ug of antibody.

Scientific Data Images for Lightning-Link (R) R-PE Antibody Labeling Kit

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0010]

Flow Cytometry: Lightning-Link R-PE Antibody Labeling Kit [703-0010] - Direct labeling with Lightning Link CD45RC-PE

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0010]

Flow Cytometry: Lightning-Link R-PE Antibody Labeling Kit [703-0010] - Indirect labeling with CD45RC supernatant

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0010]

Flow Cytometry: Lightning-Link R-PE Antibody Labeling Kit [703-0010] - Indirect labeling with purified CD45RC antibody

Kit Contents for Lightning-Link (R) R-PE Antibody Labeling Kit

1 or 3 or 5 glass vial(s) of Lightning-Link mix
1 vial of LL-Modifier reagent
1 vial of LL-Quencher reagent

Formulation, Preparation, and Storage

Concentration

Concentration is not relevant for this product. Please see the protocols for proper use of this product.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -20C.

Background: Lightning-Link (R) R-PE

Easy R-PE labeling of your antibody with 30 seconds hands-on time. R-PE (R-Phycoerythrin) is a large 240kDa red phycobiliprotein isolated from red algae. R-PE has a strong absorption peak at 565nm and a secondary absorption peak at 492nm. The R-PE protein is commonly used for fluorescent immunolabeling, particularly in applications involving fluorescent-activated cell sorting. The Lightning-Link labeling system represents a quantum leap forward in antibody labeling technology. It allows the researcher to make labeled antibodies with minimal hands-on time - less than 30 seconds. Lightning-Link simplifies immunoassay techniques, such as western blotting, ELISA and immunohistochemistry, by eliminating secondary reagents and cutting the number of incubation and wash steps. Despite its simplicity, Lightning-Link is a very sophisticated conjugation system in which the antibody is directionally coupled to the label (and not to itself) in a controlled fashion, creating high quality labeled antibodies and proteins. As the procedure is not interrupted by desalting steps, trial labels can be prepared with microgram quantities of protein and then scaled up with ease. Lightning-Link eliminates problems associated with scale up; hands-on time is essentially independent of process scale. Consistency from batch to batch is easy to achieve, and recoveries approach 100%.

Additional Lightning-Link (R) R-PE Products

Product Documents for Lightning-Link (R) R-PE Antibody Labeling Kit

Product Datasheets

Download Product Datasheets

Protocols & Manuals

Download Manual - Lightning-Link (R) R-PE Antibody Labeling Kit

COA

SDS

Download SDS

Product Specific Notices for Lightning-Link (R) R-PE Antibody Labeling Kit

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. Kits are guaranteed for 6 months from date of receipt.

Citations
Reviews

Protocols
FAQs

Lightning-Link (R) R-PE Antibody Labeling Kit

Novus Biologicals, part of Bio-Techne | Catalog # 703-0010

Key Product Details

Applications

Kit Type

Conjugate

Product Summary for Lightning-Link (R) R-PE Antibody Labeling Kit

Product Specifications

Application Notes

Scientific Data Images for Lightning-Link (R) R-PE Antibody Labeling Kit

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0010]

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0010]

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0010]

Kit Contents for Lightning-Link (R) R-PE Antibody Labeling Kit

Formulation, Preparation, and Storage

Concentration

Shipping

Storage

Background: Lightning-Link (R) R-PE

Additional Lightning-Link (R) R-PE Products

Product Documents for Lightning-Link (R) R-PE Antibody Labeling Kit

Product Datasheets

Protocols & Manuals

COA

Certificate of Analysis

SDS

Product Specific Notices for Lightning-Link (R) R-PE Antibody Labeling Kit

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