Alk. Phos Conjugation Kit

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LYNX Rapid Alkaline Phosphatase Antibody Conjugation Kit

Product Type
Conjugation Kit
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
LNK011AP CJ 1 Conjugation For 100µg Antibody
LNK014AP CJ 3 Conjugations For 10µg Antibody
LNK012AP CJ 3 Conjugations For 100µg Antibody
LNK013AP CJ 1 Conjugation For 1mg Antibody
LYNX Rapid Alkaline Phosphatase Antibody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing Alkaline Phosphatase (AP) label to an antibody or protein. Activation of proprietary reagents within the antibody-label solution results in directional covalent bonding of AP 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: 1 Conjugation For 100µg Antibody
1 Vial of 100ug LYNX lyophilized AP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 3 Conjugations For 10µg Antibody
3 Vials of 10ug LYNX lyophilized AP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 3 Conjugations For 100µg Antibody
3 Vials of 100ug LYNX lyophilized AP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 1 Conjugation For 1mg Antibody
1 Vial of 1mg LYNX lyophilized AP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Preparing The Antibody
Pack Size: 1 Conjugation For 100µg Antibody, 3 Conjugations For 100µg 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.

The molar ratio of antibody: AP should be 1:1, i.e. 100ug antibody to every 100ug AP. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 100ul and a maximum antibody amount of 100ug. 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.

* Phosphate may inhibit alkaline phosphatase activity. This kit can be used with antibodies prepared in phosphate buffer as long as the buffer will be washed away during an immunoassay procedure. Otherwise we recommend using a non-phosphate containing buffer for labeling antibodies with alkaline phosphatase.
Pack Size: 3 Conjugations For 10µg 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.

The molar ratio of antibody: AP should be 1:1, i.e. 10ug antibody to every 10ug AP. 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.

* Phosphate may inhibit alkaline phosphatase activity. This kit can be used with antibodies prepared in phosphate buffer as long as the buffer will be washed away during an immunoassay procedure. Otherwise we recommend using a non-phosphate containing buffer for labeling antibodies with alkaline phosphatase.
Pack Size: 1 Conjugation For 1mg 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.

The molar ratio of antibody: AP should be 1:1, i.e. 1mg antibody to every 1mg AP. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 1ml and a maximum antibody amount of 1mg. 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.

* Phosphate may inhibit alkaline phosphatase activity. This kit can be used with antibodies prepared in phosphate buffer as long as the buffer will be washed away during an immunoassay procedure. Otherwise we recommend using a non-phosphate containing buffer for labeling antibodies with alkaline phosphatase.

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 Alk. Phos 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 at room temperature (20-25oC) for 3 hours, or overnight if preferred.

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 Alk. Phos Conjugation Kit

  1. Pérez Ruiz, E. et al. (2016) Evaluation of different strategies for magnetic particle functionalization with DNA aptamers.
    N Biotechnol. Jun 15. pii: S1871-6784(16)32260-9. [Epub ahead of print]