Biotin Labeling Kit - NH2 (for 1mg)
Protein Labeling Kit
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Product codeLK55 Biotin Labeling Kit - NH2 (for 1mg)
| Unit size | Price | Item Code |
|---|---|---|
| 1 sample | LK55-10 |
| 1 sample | ・NH2-Reactive Biotin ・WS Buffer ・Reaction Buffer ・Filtration Tube ・15 ml Tube (for counterbalance) |
1 tube 13 ml x 1 1.2 ml x 1 1 tube 1 tube |
|---|
Description
Biotin Labeling Kit-NH2 is mainly used for the preparation of biotin-labeled IgG for enzyme immunoassay (EIA). NH2-reactive biotin, a component of this kit, has succinimidyl groups (NHS) that react with amino groups on proteins or other molecules (Fig. 1). This kit contains all the reagents necessary for the labeling. The labeling process is very simple. Just add the NH2- reactive Biotin to IgG solution and incubate at 37oC for 10 minutes. An average of 5 to 8 biotin molecules conjugate to each IgG molecule. The number of biotin molecules per protein can be determined by HABA assay. Excess biotin molecules can be removed by a filtration tube.
Fig. 1 IgG labeling reaction with NH2-reactive biotin
Precaution:
♦ The molecular weight of the protein to be labeled with this kit should be greater than 50,000.
♦ IgG or biotin-conjugated IgG is always on the membrane of the filtration tube during the labeling process.
♦ If the IgG solution contains other proteins with molecular weights larger than 10,000, such as BSA or gelatin, purify the IgG solution before labeling biotin with this kit.
♦ If the IgG solution contains small insoluble materials, centrifuge the solution and use the supernatant for the labeling.
Easily Switch Fluorescence wavelength on your primary antibody
| Developer | Dojindo Molecular Technologies, Inc |
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Manual
References
References
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Q & A
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Q
Can I use this kit for other proteins?
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A
Yes, if the molecular weight is greater than 50,000.
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Q
Do I have to use a Filtration tube prior to labeling the protein?
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A
If the protein solution does not contain small molecules with amino groups and the concentration of the protein is 10 mg per ml or about 70 μM, there is no need to use the Filtration tube. Just mix 10 μl of the sample solution with 90 μl of Reaction buffer and add the mixture to a vial of NH2-reactive Biotin. After the reaction, transfer all of the reaction mixture to a Filtration tube, and then follow the protocol starting at step 6.
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Q
Do I have to use WS buffer to store the biotin-labeled protein?
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A
You don’t have to use WS buffer. You can choose any kind of buffer according to your experiment.
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Q
My sample contains small insoluble material. What should I do?
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A
Spin the sample and use the supernatant for the labeling.
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Q
How long is the biotin-labeled protein stable?
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A
If you store the biotin-labeled protein at 0-5ºC, it is stable for 2 months. For longer storage, add 100% volume of glycerol, aliquot, and store at -20ºC (if the protein can be frozen). However, please note that stability depends on the protein itself.
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Q
What is the minimum amount of IgG that can be labeled by this kit?
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A
The minimum amount is 10 μg IgG; simply follow the protocol. The labeling ratio remains the same for 10 μg to 100 μg of IgG.
Handling and storage condition
| 0-5°C, Protect from moisture |
