Biotin (Type 1) Conjugation Kit
LYNX Rapid Plus Biotin (Type 1) Antibody Conjugation Kit (LNK262B) used to biotinylate human myelin basic protein prior to in vitro MBP binding assays wirh human B cells assessed by flow cytometry.
Image caption:
Serum complement promotes the binding of MBP to normal B cells.
PBMCs from healthy donors were incubated for 30 min with or without 30 μg/ml biotinylated MBP in medium containing normal serum (30% v/v), or in pure medium. (A) Histogram plot depicting MFI values of MBP binding to B cells in one representative healthy donor. B) The binding of MBP to B cells from 7 healthy donors is shown, expressed as percentage MBP-positive B cells. C) Before addition to the culture media, serum was treated in one of three ways: heat-inactivated (h.i.) by heating to 56°C for 30 min, or supplemented with EDTA or sodium polyanethole sulphonate (SPS) in different concentrations. The resulting MFI values from 5–7 healthy donors are shown. Bars and error bars represent means and SEM *p<0.05 **p<0.01 and ***p<0.001.
From: Brimnes MK, Hansen BE, Nielsen LK, Dziegiel MH, Nielsen CH (2014)
Uptake and Presentation of Myelin Basic Protein by Normal Human B Cells.
PLoS ONE 9(11): e113388.
doi: 10.1371/journal.pone.0113388
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
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CJ ResetLYNX Rapid Plus Biotin (Type 1) Antibody Conjugation Kit
- Product Type
- Conjugation Kit
- Specificity
- Biotin (Type 1) Conjugation Kit
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| LYNX Rapid Plus Biotin (Type 1) Antibody Conjugation Kit® enables the rapid conjugation of a pre-prepared lyophilized mixture containing Biotin label to an antibody or protein in minutes. Activation of proprietary reagents within the antibody-label solution results in the coupling of the antibody to Biotin. The LYNX Rapid Plus 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 and no requirement for desalting or dialysis. This kit has been optimized for the additional use of a streptavidin labelled reagent. For customers intending to capture their conjugate on Streptavidin coated plates, we recommend our LYNX Rapid Plus Biotin (Type 2) Antibody Conjugation Kit instead. |
- Reagents in the Kit
- Pack Size: 3 Conjugations For 20µg Antibody3 Vials of 10μg LYNX Rapid Plus lyophilized Biotin mix.
1 Vial LYNX Modifier reagent.
1 Vial LYNX Quencher reagent.Pack Size: 3 Conjugations For 200µg Antibody3 Vials of 100μg LYNX Rapid Plus lyophilized Biotin mix.
1 Vial LYNX Modifier reagent.
1 Vial LYNX Quencher reagent.Pack Size: 1 Conjugation For 2mg Antibody1 Vial of 1mg LYNX Rapid Plus lyophilized Biotin mix
1 Vial LYNX Modifier reagent
1 Vial LYNX Quencher reagent - Preparing The Antibody
- Pack Size: 3 Conjugations For 20µg AntibodyThe 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.
Do not use buffers containing nucleophilic components (e.g primary amines), thiols (e.g. Thiomersal/Thimerosal), Merthiolate, Glycine or Proclin. These substances 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 up to 10-20 μg antibody be used in each labelling reaction. For optimal results the antibody volume should be 4-10 μl, at a concentration range between 1mg/ml and 2.5 mg/ml.Pack Size: 3 Conjugations For 200µg AntibodyThe 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.
Do not use buffers containing nucleophilic components (e.g primary amines), thiols (e.g. Thiomersal/Thimerosal), Merthiolate, Glycine or Proclin. These substances 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 up to 100-200 μg antibody be used in each labeling reaction. For optimal results the antibody volume should be 40-100 μl, at a concentration range between 1mg/ml and 2.5 mg/ml.Pack Size: 1 Conjugation For 2mg AntibodyThe 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.
Do not use buffers containing nucleophilic components (e.g primary amines), thiols (e.g. Thiomersal/Thimerosal), Merthiolate, Glycine or Proclin. These substances 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 up to 2mg antibody be used in each labeling reaction. For optimal results the antibody volume should be 400-1000μl, at a concentration range between 1mg/ml and 2.5 mg/ml. - Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
Pack Size: 3 Conjugations For 20µg Antibody, 3 Conjugations For 200µg Antibody
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 -20°C 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.
Pack Size: 1 Conjugation For 2mg Antibody
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 -20°C 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. Before you add antibody to the Lynx Rapid Plus mix, add 1 μl of Rapid Modifier reagent for each 10 μl of antibody to be labelled. Mix gently.
2. Remove the screw cap from the vial of Lynx Rapid Plus mix and pipette the antibody sample (with the added Rapid Modifier) directly onto the lyophilised material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.
3. Replace cap on the vial and leave standing for 15 minutes at room temperature (20-25 0C). Longer incubation times could have negative effects on the conjugate.
4. After 15 minutes incubation, add 1 μl of Lynx Rapid Plus Quencher reagent for every 10 μl of antibody used. The conjugate can be used after 4 minutes. The conjugates do not require purification. - 1. Before you add antibody to the Lynx Rapid Plus mix, add 1 μl of Rapid Modifier reagent for each 10 μl of antibody to be labelled. Mix gently.
2. Remove the screw cap from the vial of Lynx Rapid Plus mix and pipette the antibody sample (with the added Rapid Modifier) directly onto the lyophilised material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.
3. Replace cap on the vial and leave standing for 15 minutes at room temperature (20-25 0C). Longer incubation times could have negative effects on the conjugate.
4. After 15 minutes incubation, add 1 μl of Lynx Rapid Plus Quencher reagent for every 10 μl of antibody used. The conjugate can be used after 4 minutes. The conjugates do not require purification. - 1. Before you add antibody to the Lynx Rapid Plus mix, add 1 μl of Rapid Modifier reagent for each 10 μl of antibody to be labelled. Mix gently.
2. Remove the screw cap from the vial of Lynx Rapid Plus mix and pipette the antibody sample (with the added Rapid Modifier) directly onto the lyophilised material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.
3. Replace cap on the vial and leave standing for 15 minutes at room temperature (20-25 0C). Longer incubation times could have negative effects on the conjugate.
4. After 15 minutes incubation, add 1 μl of Lynx Rapid Plus Quencher reagent for every 10 μl of antibody used. The conjugate can be used after 4 minutes. The conjugates do not require purification.
References for Biotin (Type 1) Conjugation Kit
-
Brimnes, M.K. et al. (2014) Uptake and presentation of myelin basic protein by normal human B cells.
PLoS One. 9 (11): e113388.
- Licensed Use
- Bio-Rad LYNX Rapid Plus Conjugation kits are offered for research purposes alone, and are not intended for human, therapeutic or diagnostic use. The purchase of this conjugation kit conveys to the buyer (whether the buyer is a not-for-profit, academic or for-profit entity) the non-transferable right to use the amount of product purchased and the components of the product for in-house research. The buyer shall not sell or otherwise transfer this product, its components, or materials prepared therefrom to any third party. The buyer shall not use this product or its components for commercial purposes. For the avoidance of doubt, ‘commercial purposes’ means any activity by a party for consideration and includes, without limitation, use of the product or its components (I) in the manufacturing of conjugated materials (e.g. labeled antibodies), (ii) to provide a service, information or data, (iii) for therapeutic, diagnostic or prophylactic purposes, or (iv) for repackaging/resale, whether or not such product or its components are resold for use in research. The use of this product by the buyer constitutes agreement with the terms of this limited use label license for LYNX products.
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