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HRP Conjugation Kit

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LYNX Rapid HRP Antibody Conjugation Kit thumbnail image 1 LYNX Rapid HRP Antibody Conjugation Kit thumbnail image 2
LYNX Rapid HRP Antibody Conjugation Kit gallery image 1
Published customer image:
LYNX Rapid HRP Antibody Conjugation Kit (LNK001P) used to conjugate detection antibody reagent prior to use in an anti Drug Antibody ELISA.
Image caption:
Pembrolizumab drug and anti-drug antibody (ADA) levels in a clinical cohort. Data generated from patients with melanoma treated with pembrolizumab (n=41). (A) Pembrolizumab drug level at baseline (B/L), just prior to Cycle 2 (T1) and last available timepoint (T2; Median Cycle 7 or 21 weeks IQR Cycle 4-13). (B) pembrolizumab ADA were detectable at B/L, T1 and T2. Patients with an ADA at T1 had higher pembrolizumab drug levels (*p<0.05).

From: Sasson SC, Wilkins LE, Watson RA, Jolly C, Brain O, Klenerman P, Olsson-Brown A, Fairfax BP.
Identification of neutralising pembrolizumab anti-drug antibodies in patients with melanoma.
Sci Rep. 2021 Sep 28;11(1):19253.
doi: 10.1038/s41598-021-98700-7..
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
LYNX Rapid HRP Antibody Conjugation Kit gallery image 2
Published customer image:
LYNX Rapid HRP Antibody Conjugation Kit (LNK001P) used to conjugate detection antibody reagent prior to use in an anti Drug Antibody ELISA.
Image caption:
Pembrolizumab drug and anti-drug antibody (ADA) levels in 3 patients with results suggestive of a neutralising ADA. 3/41 patients (7%) with results suggestive of a neutralising ADA are shown. All 3 patients had falling pembrolizumab drug levels and rising ADA between Cycle 2 (T1) and last available time-point (T2; Median Cycle 7 or 21 weeks IQR Cycle 4-13).

From: Sasson SC, Wilkins LE, Watson RA, Jolly C, Brain O, Klenerman P, Olsson-Brown A, Fairfax BP.
Identification of neutralising pembrolizumab anti-drug antibodies in patients with melanoma.
Sci Rep. 2021 Sep 28;11(1):19253.
doi: 10.1038/s41598-021-98700-7..
This image is from an open access article distributed under terms of a Creative Commons Attribution License.

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LYNX Rapid HRP Antibody Conjugation Kit

Product Type
Conjugation Kit
Specificity
HRP Conjugation Kit

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Product Code Applications Pack Size List Price Your Price Qty
LNK001P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 1 Conjugation For 400µg Antibody
List Price Your Price
LNK002P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 3 Conjugations For 400µg Antibody
List Price Your Price
LNK003P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 1 Conjugation For 4mg Antibody
List Price Your Price
LNK004P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 5 Conjugations For 4mg Antibody
List Price Your Price
LNK005P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 1 Conjugation For 20mg Antibody
List Price Your Price
LNK006P
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
CJ 3 Conjugations For 40µg Antibody
List Price Your Price

LYNX Rapid HRP Antibody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing Horseradish peroxidase (HRP) label to an antibody or protein. Activation of proprietary reagents within the antibody-label solution results in directional covalent bonding of HRP 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
Pack Size: 1 Conjugation For 400µg Antibody
1 Vial of 100μg LYNX lyophilized HRP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 3 Conjugations For 400µg Antibody
3 Vials of 100ug LYNX lyophilized HRP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 1 Conjugation For 4mg Antibody
1 Vial of 1mg LYNX lyophilized HRP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 5 Conjugations For 4mg Antibody
5 Vials of 1mg LYNX lyophilized HRP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 1 Conjugation For 20mg Antibody
1 Vial of 5mg LYNX lyophilized HRP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Pack Size: 3 Conjugations For 40µg Antibody
3 Vials of 10μg LYNX lyophilized HRP mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent
Preparing The Antibody
Pack Size: 1 Conjugation For 400µg 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. EDTA and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Sodium azide is an irreversible inhibitor of HRP and therefore should be avoided.

The amount of antibody used for labeling ideally should correspond to molar ratios between 1:4 and 1:1 Ab to HRP. Taking account of the molecular weights (160,000 versus 40,000), this means for that for 100μg HRP you need to add between 100-400μg of antibody. For optimal results the antibody volume should be up to 100μl, at a concentration range of 0.5-5.0mg/ml.
Pack Size: 3 Conjugations For 400µg 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. EDTA and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Sodium azide is an irreversible inhibitor of HRP and therefore should be avoided.

The amount of antibody used for labeling ideally should correspond to molar ratios between 1:4 and 1:1 Ab to HRP. Taking account of the molecular weights (160,000 versus 40,000), this means for that for 100μg HRP you need to add between 100-400μg of antibody. For optimal results the antibody volume should be up to 100μl, at a concentration range of 0.5-5.0mg/ml.
Pack Size: 1 Conjugation For 4mg 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. EDTA and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Sodium azide is an irreversible inhibitor of HRP and therefore should be avoided.

The amount of antibody used for labeling ideally should correspond to molar ratios between 1:4 and 1:1 Ab to HRP. Taking account of the molecular weights (160,000 versus 40,000), this means for that for 1mg HRP you need to add between 1-4mg of antibody.. For optimal results the antibody volume should be up to 1ml, at a concentration range of 0.5-5.0mg/ml.
Pack Size: 5 Conjugations For 4mg 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. EDTA and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Sodium azide is an irreversible inhibitor of HRP and therefore should be avoided.

The amount of antibody used for labeling ideally should correspond to molar ratios between 1:4 and 1:1 Ab to HRP. Taking account of the molecular weights (160,000 versus 40,000), this means for that for 4mg HRP you need to add between 1-4mg of antibody. For optimal results the antibody volume should be up to 1ml, at a concentration range of 0.5-5.0mg/ml.
Pack Size: 1 Conjugation For 20mg 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. EDTA and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Sodium azide is an irreversible inhibitor of HRP and therefore should be avoided.

The amount of antibody used for labeling ideally should correspond to molar ratios between 1:4 and 1:1 Ab to HRP. Taking account of the molecular weights (160,000 versus 40,000), this means for that for 5mg HRP you need to add between 5-20mg of antibody. For optimal results the antibody volume should be up to 5ml, at a concentration range of 0.5-5.0mg/ml.
Pack Size: 3 Conjugations For 40µg 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. EDTA and common non-buffering salts and sugars have little or no effect on conjugation efficiency.

Sodium azide is an irreversible inhibitor of HRP and therefore should be avoided.

The amount of antibody used for labeling ideally should correspond to molar ratios between 1:4 and 1:1 Ab to HRP. Taking account of the molecular weights (160,000 versus 40,000), this means for that for 10μg HRP you need to add between 10-40μg of antibody. For optimal results the antibody volume should be up to 10μl, at a concentration range of 0.5-5.0mg/ml.
Regulatory
For research purposes only
Guarantee
12 months from date of despatch

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.
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 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 at room temperature (20-25°C) 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.

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.

1. To the antibody sample add 1μl 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 1μl of Quencher reagent for every 10μl of antibody used. Leave to stand for 30 minutes before 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 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.

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 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.

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 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.

References for HRP Conjugation Kit

  1. Bondzio, A. et al. (2011) Identification of differentially expressed proteins in ruminal epithelium in response to a concentrate-supplemented diet.
    Am J Physiol Gastrointest Liver Physiol. 301 (2): G260-8.
  2. Lichtmannegger, J. et al. (2016) Methanobactin reverses acute liver failure in a rat model of Wilson disease.
    J Clin Invest. 126 (7): 2721-35.
  3. Sasson, S.C. et al. (2021) Identification of neutralising pembrolizumab anti-drug antibodies in patients with melanoma.
    Sci Rep. 11 (1): 19253.
  4. Rosadas, C. et al. (2022) Detection and quantification of antibody to SARS CoV 2 receptor binding domain provides enhanced sensitivity, specificity and utility.
    J Virol Methods. 302: 114475.
  5. Khan, M. et al. (2022) Simple, sensitive, specific self-sampling assay secures SARS-CoV-2 antibody signals in sero-prevalence and post-vaccine studies.
    Sci Rep. 12 (1): 1885.

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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