Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) – Lightning-Link®

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Overview

Product name

Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) – Lightning-Link®

Product overview

Biotinylation Kit / Biotin Conjugation Kit (Type A) (ab201795) uses a simple and quick process for biotinylation / biotin labeling of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.


To conjugate an antibody to Biotin using this kit:
– add modifier to antibody and incubate for 15 mins
– add quencher and incubate for 5 mins
The biotin conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.


Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to Biotin.


The Type A Biotinylation Kit / Biotin Conjugation Kit has been optimized to produce conjugates for assays in which a streptavidin-labeled detection reagent will be used.


Use the Type B Biotinylation Kit / Biotin Conjugation Kit ab201796 for assays in which the biotinylated protein is captured by streptavidin immobilized on a surface (e.g. plates, nitrocellulose, magnetic beads etc).


Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.

Notes

This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Rapid Biotin Type A Labeling Kit. 370-0005 is the same as the 100 µg size. 370-0010 is the same as the 3 x 100 ug size. 370-0030 is the same as the 3 x 10 ug size. 370-0015 is the same as the 1 mg size.

Amount and volume of antibody for conjugation to Biotin

 Kit sizeRecommended 
amount of antibody
1 
Maximum 
amount of antibody
Maximum antibody 
volume2
3 x 10 µg3 x 10 µg 3 x 20 µg3 x 10 µL
100 µg1 x 100 µg1 x 200 µg1 x 100 µL
3 x 100 µg3 x 100 µg 3 x 200 µg3 x 100 µL
1 x 1 mg1 x 1 mg1 x 2 mg1 x 1 mL

Using the maximum amount of antibody may result in less labelling per antibody. 

2 Ideal antibody concentration is 1mg/ml. 0.5 – 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2mg/ml or < 0.5 mg/ml should be diluted /concentrated.

 

Buffer Requirements for Conjugation

Buffer should be pH 6.5-8.5.

Compatible buffer constituents 
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.

50mM / 0.6% Tris10.1% BSA250% glycerol
0.1% sodium azidePBSPotassium phosphate
Sodium chlorideHEPESSucrose
Sodium citrateEDTATrehalose

1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 BSA can also interfere with the use of the conjugated antibody in tissue staining.

Incompatible buffer constituents

ThiomerosalProclinGlycine
ArginineGlutathioneDTT

If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.

Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.

Storing and handling conjugation kits

Lyophilized Lightning-Link® components are hygroscopic.

Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.

Properties

Storage instructions

Store at -20°C. Please refer to protocols.
Components1 mg100 µg3 x 100 µg3 x 10 µg
ab274076 – Biotin (Type A) Conjugation Mix1 x 1mg1 x 100µg3 x 100µg3 x 10µg
ab273994 – Modifier reagent1 x 200µl1 x 200µl1 x 200µl1 x 200µl
ab273995 – Quencher reagent1 x 200µl1 x 200µl1 x 200µl1 x 200µl

Research areas

  • Tags & Cell Markers
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  • Epitope Tags
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  • Conjugates
  • Signal Transduction
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  • Metabolism
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  • Vitamins / Minerals
  • Kits/ Lysates/ Other
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  • Kits
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  • Conjugation Kits
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  • Biotin

Protocols