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RPE-Cy5 Conjugation Kit

LYNX Rapid RPE-Cy5 Antibody Conjugation Kit

Product Type
Conjugation Kit
Specificity
RPE-Cy5 Conjugation Kit

Product Code Applications Pack Size List Price Your Price Qty
LNK084C
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 3 Conjugations For 10µg Antibody loader
List Price Your Price
loader
LNK082C
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 3 Conjugations For 60µg Antibody loader
List Price Your Price
loader

LYNX Rapid RPE-Cy5 Antibody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing R-Phycoerythrin (RPE)-Cy5 label to an antibody or protein. Activation of proprietary reagents within the antibody-label solution results in directional covalent bonding of RPE-Cy5 to the antibody.

The LYNX Rapid Conjugation kit® can be used to label small quantities of antibody/protein at near neutral pH, allowing a high conjugation efficiency with 100% antibody recovery.

Reagents In The Kit
LNK084C: 3 Vials of 10μg LYNX lyophilized RPE-Cy5 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent.
LNK082C: 3 Vials of 100μg LYNX lyophilized RPE-Cy5 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Preparing The Antibody
LNK084C: The following buffer solutions are recommended for preparing the antibody:

10-50mM amine-free buffer (e.g HEPES, MES, MOPS and phosphate) pH range 6.5-8.5, although moderate concentrations of Tris buffer (<20mM) may be tolerated.

If possible, avoid buffers containing nucleophilic components such as primary amines and thiols (e.g. thiomersal/thimerosal) since they may react with LYNX chemicals. Azide (0.02-0.1%), EDTA, up to 50% Glycerol and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Due to the large size of the RPE-Cy5 label, the quantity of RPE-tandem dye is in slight molar excess. Approximately 10ug of IgG will give a 1:1 molar ratio of antibody:RPE. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 10ul and a maximum antibody amount of 10ug. Antibody at a concentration of greater than 1mg/ml requires dilution. Antibody below 1mg/ml can still be used as long as the maximum volume is not exceeded. Using less than the recommended amount of antibody may result in unbound label, but this will be removed during subsequent application wash steps. Antibody below 0.5mg/ml should be concentrated before use with the kit.
LNK082C: The following buffer solutions are recommended for preparing the antibody:

10-50mM amine-free buffer (e.g HEPES, MES, MOPS and phosphate) pH range 6.5-8.5, although moderate concentrations of Tris buffer (<20mM) may be tolerated.

If possible, avoid buffers containing nucleophilic components such as primary amines and thiols (e.g. thiomersal/thimerosal) since they may react with LYNX chemicals. Azide (0.02-0.1%), EDTA, up to 50% Glycerol and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Due to the large size of the RPE-Cy5 label, it is recommended that 50-60ug of antibody be used for every 100μg RPE-Cy5, to ensure a slight RPE-Cy5 molar excess (50μg antibody gives a 1:1 Ab:RPE-Cy5 molar ratio). For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 60ul and a maximum antibody amount of 60ug. Antibody at a concentration of greater than 1mg/ml requires dilution. Antibody below 1mg/ml can still be used as long as the maximum volume is not exceeded. Using less than the recommended amount of antibody may result in unbound label, but this will be removed during subsequent application wash steps. Antibody below 0.5mg/ml should be concentrated before use with the kit.
Regulatory
For research purposes only
Guarantee
12 months from date of despatch
Acknowledgements
This product or portions thereof is manufactured under license from Carnegie Mellon University under U.S. Patent Number 5,268,486 and related patents. Cy and CyDye are trademarks of GE Healthcare Limited.

This kit contains lyophilized hygroscopic components that are moisture-sensitive. This kit is shipped under ambient conditions with silica packets to avoid exposure to moisture. On receipt, Bio-Rad recommend that the kit is stored at -20oC and protected from moisture. Storage in frost-free freezers is not recommended.This product should be stored undiluted. Avoid repeated freezing and thawing. Before opening, allow the components to reach room temperature to minimize condensation.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Conjugation

We recommend that for each conjugation the user determines the best antibody:conjugate ratio.
Instructions For Use
1. To the antibody sample add 1μl of the Modifier reagent for every 10μl of antibody and mix gently.

2. Pipette the mixed antibody-modifier sample directly onto the LYNX lyophilized mix and gently pipette up and down twice to resuspend.

3. Replace cap onto vial and incubate in the dark at room temperature (20-25oC) for 3 hours, or overnight if preferred.

4. After incubation, add 1μl of Quencher reagent for every 10μl of antibody used. Leave to stand for 30 minutes before use.

1. To the antibody sample add 1μl of the Modifier reagent for every 10μl of antibody and mix gently.

2. Pipette the mixed antibody-modifier sample directly onto the LYNX lyophilized mix and gently pipette up and down twice to resuspend.

3. Replace cap onto vial and incubate in the dark at room temperature (20-25oC) for 3 hours, or overnight if preferred.

4. After incubation, add 1μl of Quencher reagent for every 10μl of antibody used. Leave to stand for 30 minutes before use.

Licensed Use
These products and the methodology of conjugation are patent protected under United Kingdom patent number 2446088 and associated international patent applications. The purchase of this product conveys to the buyer the limited, non exclusive non-transferable right (without the right to resell repackage or further sublicense) under these patents to use the product to make conjugates for research and development purposes only. The purchaser cannot sell or otherwise transfer this product, or its components, or materials or data made using this product, or its components to a third party. Further information on purchasing licenses for diagnostic and other uses may be obtained by contacting Bio-Rad, at. Endeavour House, Langford Business Park, Langford Lane, Kidlington, Oxon. OX5 1GE UNITED KINGDOM. Tel: +44 1865 852 700. E-mail: antibodies@bio-rad.com

LNK084C

149787 162350 167137

LNK082C

148735 154887 159326 161248 164668 166263

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