Product Catalog

Product Image: 
BIS-300 positive and negative control protein kit
SKU:
BIS - 300
$150.00

The BIS-300 Kit contains a fully biotinylated MBP-AviTagTM fusion protein standard and an unbiotinylated MBP-AviTagTM fusion protein that may be used as an "extent-of-biotinylation" comparison for BirA biotin-protein ligase reactions, for SDS-PAGE gel analysis or in Western blot analysis.

Kit supplied with the following:

• One (1) vial containing 50μL of fully biotinylated MBP-AviTagTM fusion protein at 1mg/mL (50μg).

• One (1) vial containing 100μL of unbiotinylated MBP-AviTagTM fusion protein at 1mg/mL (100μg).

By comparing a known amount of a 100% biotinylated AviTagTM fusion protein as a standard against a known amount of BirA-reacted protein-of-interest in an ELISA-type, multi-well dilution format (called BRTA for Biotinylation Reaction Titration Assay) it is possible to assess the extent of biotinylation achieved in your own BirA biotinylation reactions in a highly accurate and reproducible manner.

In addition, a mass shift between biotinylated and unbiotinylated fusion proteins visible on both SDS-PAGE and Western blot can be used to subjectively determine the amount of biotinylation that has occurred in a reaction by the observation of differences in band density in a mixed population of biotinylated vs. nonbiotinylated substrate.

The BIS-300 Kit contains the fully biotinylated standard used for the BRTA protocol and an unbiotinylated AviTag’d protein to react in a parallel to your BirA/protein biotinylation reaction as a positive control. A link to the BRTA protocol can be found here.

  • What is the amino acid sequence for your MBP-AviTag fusion protein?

    The sequence is provided below.

    MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLA

    EITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTW

    PLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNI

    DTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPR

    IAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSGSLSTPPTPSPSTPPTGLNDIFEA

    QKIEWHE

    Color legend: MBP-IgA hinge linker-AviTag