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 would like to know that the alkaline phosphatase used in this labeling kit is produced in E. coli or bovine.

The Alkaline Phosphatase that we use is from a bovine source.

What is its pH optimum?

The Alkaline Phosphatase that we use is from a bovine source. We believe the optimum pH to be around 9.6-9.8, so if you are looking to conduct an ELISA post conjugation, you may wish to use a substrate mix at this pH. It may be worth noting however that this pH is outside of the range for conjugation using the Lightning-Link kits, so you would need to modify the pH after the conjugation reaction is complete.

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.

Lightning-Link (R) Alkaline Phosphatase Antibody Labeling Kit

Name: Lightning-Link (R) Alkaline Phosphatase Antibody Labeling Kit
Brand: Novus Biologicals
SKU: 702-0010
Rating: 5 (1 reviews)

Novus Biologicals, part of Bio-Techne | Catalog # 702-0010

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Product Summary
Product Specifications
Scientific Data
Kit Contents
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Specific Notices

Key Product Details

Applications

ELISA

Kit Type

Antibody Labeling Kit

Conjugate

Alkaline Phosphatase

Product Summary for Lightning-Link (R) Alkaline Phosphatase 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 Alkaline Phosphatase antibody labeling kit enables the direct conjugation of Alkaline Phosphatase to any biomolecule with an available amine group. The researcher simply pipettes their antibody or other biomolecule into the vial of Lightning-Link label 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

Alkaline phosphatase (AP) is a hydrolyase enzyme that is frequently conjugated to antibodies for use in immunoassays. PNPP is a popular colorimetric substrate for AP; the product of the reaction, p-nitrophenol, can be read at 405nM. Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate; MUP) may also be employed.

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 100 ug of antibody.

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

ELISA: Lightning-Link Alkaline Phosphatase Antibody Labeling Kit [702-0010]

Kit Contents for Lightning-Link (R) Alkaline Phosphatase Antibody Labeling Kit

1 or 3 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) Alkaline Phosphatase

Easy Alkaline Phosphatase labeling of your antibody with 30 seconds hands-on time. The Lightning-Link conjugation system represents a quantum leap forward in conjugation technology. It allows you to make antibody conjugates 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 conjugates. As the procedure is not interrupted by desalting steps, trial conjugates 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) Alkaline Phosphatase Products

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

Product Datasheets

Download Product Datasheets

Protocols & Manuals

Download Manual - Lightning-Link (R) Alkaline Phosphatase Antibody Labeling Kit

COA

SDS

Download SDS

Product Specific Notices for Lightning-Link (R) Alkaline Phosphatase 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.

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