RPE-Cy5.5 Conjugation Kit
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
- 3 Vials of 10μg LYNX lyophilized RPE-Cy5.5 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
- Preparing The Antibody
- 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-Cy5.5 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.
- Store kit at -20oC only.
Newly-conjugated antibody can be stored at 4oC. For long term storage however, the addition of a preservative is recommended.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted.
Avoid repeated freezing and thawing.
- 12 months from date of despatch
- 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.
- 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: firstname.lastname@example.org
- For research purposes only
Applications of RPE-Cy5.5 Conjugation Kit
|Application Name||Verified||Min Dilution||Max Dilution|
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.