RPE-Cy7 Conjugation Kit
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LYNX Rapid RPE-Cy7 Antibody Conjugation Kit
- Product Type
- Conjugation Kit
- Specificity
- RPE-Cy7 Conjugation Kit
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| LYNX Rapid RPE-Cy7 Antibody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing R-Phycoerythrin (RPE)-Cy7 label to an antibody or protein. Activation of proprietary reagents within the antibody-label solution results in directional covalent bonding of RPE-Cy7 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
-
LNK114PECY7: 3 Vials of 10ug LYNX lyophilized RPE-Cy7 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent -
LNK113PECY7: 1 Vial of 1mg LYNX lyophilized RPE-Cy7 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent -
LNK112PECY7: 3 Vials of 100ug LYNX lyophilized RPE-Cy7 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent -
LNK111PECY7: 1 Vial of 100ug LYNX lyophilized RPE-Cy7 mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent - Preparing The Antibody
-
LNK114PECY7: 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-Cy7 label the quantity of RPE-tandem dye is in slight molar excess. 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. -
LNK113PECY7: 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-Cy7 label, it is recommended that 50-60ug of antibody be used for every 100ug RPE-Cy7, to ensure a slight RPE-Cy7 molar excess (50ug antibody gives a 1:1 Ab:RPE-Cy7 molar ratio). 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. -
LNK112PECY7, LNK111PECY7: 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-Cy7 label, it is recommended that 50-60ug of antibody be used for every 100ug RPE-Cy7, to ensure a slight RPE-Cy7 molar excess (50ug antibody gives a 1:1 Ab:RPE-Cy7 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.
We recommend that for each conjugation the user determines the best antibody:conjugate ratio.
| 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. - 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. - 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. - 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.
References for RPE-Cy7 Conjugation Kit
-
Gawronska-Kozak, B. et al. (2021) Dermal White Adipose Tissue (dWAT) Is Regulated by Foxn1 and Hif-1α during the Early Phase of Skin Wound Healing.
Int J Mol Sci. 23 (1)Dec 27 [Epub ahead of print]. -
Haach, V. et al. (2023) A polyvalent virosomal influenza vaccine induces broad cellular and humoral immunity in pigs.
Virol J. 20 (1): 181. -
Rotolo, A. et al. (2023) Unedited allogeneic iNKT cells show extended persistence in MHC-mismatched canine recipients.
Cell Rep Med. 4 (10): 101241.
- 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
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