This product is typically not processed using animal component materials. This BirA Ligase is recombinant from E. coli. However, the sources of these materials may change from lot to lot and is reported on the Certificate of Origin. To check the source of a current lot, we kindly ask you to navigate to the link https://www.sigmaaldrich.com/techservice, click on "Product Documentation" under the Products Section with all the required information so that a member of our team can reach out to you to assist further. Thank you.
CS0008
Enzymatic Protein Biotinylation Kit
for biotinylation of up to 20 mg target protein
Synonym(s):
Biotin labeling kit
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About This Item
NACRES:
NA.84
UNSPSC Code:
12161503
Pricing and availability is not currently available.
General description
Biotinylated proteins are used in a variety of applications, such as binding assays, biopanning, and protein purification. Although chemical biotinylation methods are common, they present several challenges, including partial labeling and high batch to-batch variation, because of the random nature of these methods. In addition, the lack of control of the biotinylation site might affect the biotinylated protein′s activity. The Enzymatic Protein Biotinylation Kit allows direct and specific biotinylation using a specific biotin ligation enzyme. The kit contains all the components required for in vitro biotinylation of up to 20 mg recombinant target protein. Biotinylation is mediated by the enzyme BirA biotin ligase, which uniformly ligates biotin to lysine residues in the context of several tags, such as AviTag™ and Biotag. This kit was tested successfully under a wide range of conditions, including:
Supportive calculator (Click here to download a calculator excel file): Will calculate the reagents needed according to your experimental needs.
- Temperatures ranging from 4 °C to 30 °C
- pH from 6.0 to 9.0
- NaCl concentrations from 50 to 500 mM.
Supportive calculator (Click here to download a calculator excel file): Will calculate the reagents needed according to your experimental needs.
Legal Information
AviTag is a trademark of Avidity, LLC
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| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| storage temp. −70°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
| shipped in dry ice | shipped in wet ice | shipped in - | shipped in - |
Storage Class
10 - Combustible liquids
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Is this product Animal Component Free?
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Does the BirA ligase contain a His tag?
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The product CS0008A BirA ligase has no tags.
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Is the biotinylation kit compatible with 500 mM imidazole in the sample buffer?
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The Enzymatic Protein Biotinylation Kit has not been tested with buffers containing imidazole, including 500 mM imidazole. The kit has been evaluated under a broad range of conditions, including pH 6.0–9.0 and NaCl concentrations from 50 mM to 500 mM. Compatible buffers include MES, Tris, Bicine, and PBS. Therefore, compatibility or performance in the presence of 500 mM imidazole cannot be guaranteed. It is advisable to test the kit with the buffer, possibly at different imidazole concentrations, although optimal results are not assured.
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The gel for product #CS0008 suggests a similar activity of the kit at different pH values (6 - 9). However, in my hands, the reaction proceeds significantly faster at pH 8.3 compared to pH 6/7. Which conditions were used for the shown labeling reactions?
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The labeling reactions for this kit were performed at pH 7.
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Is the BirA protein tagged in any way (ideally FLAG) in order to remove it from the reaction mixture?
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The BirA ligase is not tagged. There is no suggested protocol for the removal of excess ligase. However, in the presence of ATP and biotin the reaction will run to completion. Any residual ligase will remain inert. The BirA ligase has a molecular weight of 33.5 kDa. Note that excess biotin is removed by dialysis. Please see the link below to review the kit Technical Bulletin:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/700/170/cs0008bul-ms.pdfHelpful?
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