The product CS0008A BirA ligase has no tags.
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$757.00
$757.00
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| shipped in dry ice | shipped in wet ice | shipped in - | shipped in dry ice |
| storage temp. −70°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level - |
General description
- 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.
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Storage Class
10 - Combustible liquids
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Does the BirA ligase contain a His tag?
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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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