Fluorescein Conjugation Kit

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LYNX Rapid Fluorescein Antibody Conjugation Kit

Product Type
Conjugation Kit
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
LNK063F CJ 3 Conjugations For 20µg Antibody
LNK061F CJ 3 Conjugations For 200µg Antibody
LNK062F CJ 1 Conjugation For 2mg Antibody
LYNX Rapid Fluorescein Antibody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing Fluorescein label to an antibody or protein. Activation of proprietary reagents within the antibody-label solution results in directional covalent bonding of Fluorescein 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.

Product Details

Reagents in the Kit
Pack Size: 3 Conjugations For 20µg Antibody, 3 Conjugations For 200µg Antibody
3 Vials LYNX lyophilized Fluorescein mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 1 Conjugation For 2mg Antibody
1 Vial LYNX lyophilized Fluorescein mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Preparing The Antibody
Pack Size: 3 Conjugations For 20µg 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 and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

It is recommended that 10-20ug antibody be used in each labelling reaction. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 10ul and a recommended antibody amount of 10ug. A maximum of 20ug of antibody can be used to obtain good quality conjugates as long as the maximum conjugation volume of 10ul is not exceeded. 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.
Pack Size: 3 Conjugations For 200µg 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 and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

It is recommended that 100-200ug antibody be used in each labelling reaction. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 100ul and a recommended antibody amount of 100ug. A maximum of 200ug of antibody can be used to obtain good quality conjugates as long as the maximum conjugation volume of 100ul is not exceeded. 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.
Pack Size: 1 Conjugation For 2mg 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 and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

It is recommended that 1-2mg antibody be used in each labelling reaction. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 1ml and a recommended antibody amount of 1mg. A maximum of 2mg of antibody can be used to obtain good quality conjugates as long as the maximum conjugation volume of 1ml is not exceeded. 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.

Storage Information

Storage
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.
Shelf Life
12 months after despatch

More Information

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
Regulatory
For research purposes only

Applications of Fluorescein Conjugation Kit

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 1ul of the Modifier reagent for every 10ul 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) overnight.

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

Product Specific References

References for Fluorescein Conjugation Kit

  1. Walker, E. et al. (2014) Microscopic Detection of Quenched Activity-Based Optical Imaging Probes Using an Antibody Detection System: Localizing Protease Activity.
    Mol Imaging Biol. 16: 608-18.
  2. Mahata, B. et al. (2014) Single-cell RNA sequencing reveals T helper cells synthesizing steroids de novo to contribute to immune homeostasis.
    Cell Rep. 7: 1130-42.