RPE Conjugation Kit

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LYNX Rapid RPE Antibody Conjugation Kit

Product Type
Conjugation Kit
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
LNK021RPE CJ 1 Conjugation For 60µg Antibody
LNK022RPE CJ 3 Conjugations For 60µg Antibody
LNK023RPE CJ 1 Conjugation For 600µg Antibody
LNK024RPE CJ 3 Conjugations For 10µg Antibody
LYNX Rapid RPE Anitbody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing R-Phycoerythrin (RPE) label to an antibody or protein. Activation of proprietary reagents within the antibody-label solution results in directional covalent bonding of RPE 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 60µg Antibody
1 Vial of 100ug LYNX lyophilized RPE mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 3 Conjugations For 60µg Antibody
3 Vials of 100ug LYNX lyophilized RPE mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 1 Conjugation For 600µg Antibody
1 Vial of 1mg LYNX lyophilized RPE mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 3 Conjugations For 10µg Antibody
3 Vials of 10ug LYNX lyophilized RPE mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Preparing The Antibody
Pack Size: 1 Conjugation For 60µg Antibody, 3 Conjugations For 60µg 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 RPE (240kDa), it is recommended that 50-60ug of antibody be used for every 100ug RPE, to ensure a slight RPE molar excess. 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.
Pack Size: 1 Conjugation For 600µg 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 RPE (240kDa), it is recommended that 50-60ug of antibody be used for every 100ug RPE, to ensure a slight RPE molar excess. For optimal results the antibody should be at a concentration of 1mg/ml, with a maximum volume of 600ul and a maximum antibody amount of 600ug. 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.
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, 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 RPE (240kDa), the quantity of RPE 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.

Storage Information

Storage
Pack Size: 1 Conjugation For 60µg Antibody, 1 Conjugation For 600µg Antibody, 3 Conjugations For 10µg Antibody
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.
Pack Size: 3 Conjugations For 60µg Antibody
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 RPE 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) 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 RPE Conjugation Kit

  1. Li, X. et al. (2010) Design of a potent CD1d-binding NKT cell ligand as a vaccine adjuvant.
    Proc Natl Acad Sci U S A. 107: 13010-5.
  2. Campbell, J.E. et al. (2010) Cellular regulation of blood coagulation: a model for venous stasis.
    Blood. 116: 6082-91.
  3. Tighe, R.M. et al. (2011) Ozone Inhalation Promotes CX3CR1-Dependent Maturation of Resident Lung Macrophages That Limit Oxidative Stress and Inflammation.
    J Immunol. 187: 4800-8.
  4. Dutertre, C.A. et al. (2008) A novel subset of NK cells expressing high levels of inhibitory FcgammaRIIB modulating antibody-dependent function.
    J Leukoc Biol. 84: 1511-20.
  5. Wielgosz, M.M. et al. (2015) Generation of a lentiviral vector producer cell clone for human Wiskott-Aldrich syndrome gene therapy
    Met Clin Dev 2, Article number: 14063
  6. Hofer, C.C. et al. (2015) Infection of Mice with Influenza A/WSN/33 (H1N1) Virus Alters Alveolar Type II Cell Phenotype.
    Am J Physiol Lung Cell Mol Physiol. ajplung.00373.2014.
  7. Welinder, C. et al. (2015) Cytokeratin 20 improves the detection of circulating tumor cells in patients with colorectal cancer.
    Cancer Lett. 358:43-6.
  8. Shive, C.L. et al. (2014) Inflammatory cytokines drive CD4+ T-cell cycling and impaired responsiveness to interleukin 7: implications for immune failure in HIV disease.
    J Infect Dis. 210: 619-29.
  9. Huang, L. et al. (2015) c-Jun gene-modified Schwann cells: upregulating multiple neurotrophic factors and promoting neurite outgrowth.
    Tissue Eng Part A. 21 (7-8): 1409-21.
  10. Hofer, S. et al. (2016) RAGE-mediated inflammation in patients with septic shock.
    J Surg Res. 202 (2): 315-27.
  11. Attatippaholkun, N. et al. (2017) Dengue Virus and Its Relation to Human Glycoprotein IIb/IIIa Revealed by Fluorescence Microscopy and Flow Cytometry.
    Viral Immunol. Sep 25 [Epub ahead of print].