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.

I want to use your product, Lightning Link R-Phycoerythrin Conjugation Kit (703-0015), to label a free amine group on a linker attached to my target. Can you give me further details on the nature of the conjugation as I wish to determine if this is applicable for my needs?

This kit is produced by a company called Innova and we distribute it for them. It operates by labeling the free amine groups, so should be suitable for your application. The key to using these kits efficiently is maintaining the proper ratios and this should work just fine.

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 accidentally placed R-PE antibobdy conjugate at -20C without glycerol, caught mistake but sample had frozen by the time I realized. What damage likely happened to conjugate and should I consider it unusable now?

Your directly conjugated antibody may still be functional. Depending on the antibody, repeated freeze thaw cycles can cause a reduction of signal or non-specific staining. Since your antibody only went through one freeze thaw cycle, it may not have been too badly damaged. If possible, I would recommend testing a small amount on a positive control or inexpensive sample.

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.

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

Name: Lightning-Link (R) R-PE Antibody Labeling Kit
Brand: Novus Biologicals
SKU: 703-0015
Rating: 5 (1 reviews)

Novus Biologicals, part of Bio-Techne | Catalog # 703-0015

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Product Summary
Product Specifications
Scientific Data
Kit Contents
Preparation & Storage
Background
Product Documents
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 can be used to label up to 600 ug of antibody, and is supplied in one vial.

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

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0015]

Flow Cytometry: Lightning-Link R-PE Antibody Labeling Kit [703-0015] - Direct labeling with Lightning Link CD45RC-PE

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0015]

Flow Cytometry: Lightning-Link R-PE Antibody Labeling Kit [703-0015] - Indirect labeling with CD45RC supernatant

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0015]

Flow Cytometry: Lightning-Link R-PE Antibody Labeling Kit [703-0015] - 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

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 # 703-0015

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 [703-0015]

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0015]

Flow Cytometry: Lightning-Link (R) R-PE Antibody Labeling Kit [703-0015]

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

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

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