In vitro Biotinylation of Avitag with Enzyme
| Avidity 主要開發及販售與分子結合相關的產品。 其專利的 AviTag™ 是可以被 BirA 酵素進行生物素鍵結反應 (Biotinylation) 的胜肽鏈 (peptide)，結合 Biotin 可與 avidin 或 streptavidin 專一性鍵結的特性，提供了理想的分子交互作用分析工具。其產品包含了in vivo, in vitro biotinylation 產品，含 AviTaq 的殖體 (vector)、Biotinylated protein、純化辨識 Avitag 的抗體…等。 |
| https://www.avidity.com/ |
| 主力產品: BirA biotin-protein ligase bulk reaction kit |
| The BirA biotin-protein ligase adds d-biotin covalently to biotin-acceptor peptides/proteins via an ATP intermediate (biotinyl 5′-adenylate) in a highly efficient and targeted manner. This kit includes 300 µg of BirA enzyme for those who require larger scale reactions or more frequent use of the BirA biotinylation reaction in their work. Please Note: We strongly recommend the use of the AviTag amino acid sequence over other Biotinylation of Peptides sequences. The BirA enzyme biotinylates the AviTag sequence at a reaction rate 2x the natural substrate (BCCP) and as much as an order of magnitude or more over the other peptide sequences in use. The AviTag™ requires smaller amounts of enzyme and shorter incubation times than other Biotinylation of Peptides sequences thus minimizing ancillary problems associated with proteases and protein instability. |
| Cell lines- in vivo biotinylationAVB101AVB101- glycerol stock CVB101- chemically competent cellsEVB101- electro-competent cellsAVB100 AVB100- glycerol stockEVB100- electro-competent cellsAVB99AVB99-glycerol stockBIP-300- positive control plasmid for induction Enzyme- in vitro biotinylationBiotin ligaseBirA500- 40ug biotin ligaseBulk BirA- 300 ug biotin ligaseLarge quantities of biotin ligaseBIS-300 positive and negative control protein kitBIO-200 biotin solutionBIO-500 biotin solution Vectors containing AviTagC-terminus AvitagpAC4AC5pAC6All threeSequencing primersN-terminus AvitagpAN4pAN5pAN6All threeSequencing primers Purified Biotinylated Proteins and support chemicalsGaussia princeps LuciferaseGluc-10Gluc-100Renilla reniformis Green Fluorescent ProteinStreptavidin-conjugated proteinsStreptavidin-Gaussia princeps LuciferaseStreptavidin-Renilla reniformis GFPCoelenterazine Purification and Identification of Avitag’d proteinsAbC- antibody against C-terminus AvitagAbCA -Abc antibody conjugated to agaroseBIS-300 positive and negative control protein kit |
產品
BirA Enzyme and buffers for in vitro biotinylation
BirA500: BirA biotin-protein ligase standard reaction kit
BirA500-RT: BirA biotin-protein ligase lyophilized reaction kit
Bulk BirA: BirA biotin-protein ligase bulk reaction kit
Bulk BirA-RT: BirA biotin-protein ligase bulk lyophilized reaction kit
BIS-300 positive and negative control protein kit
Cell lines for in vivo biotinylation
AVB101 – E.coli B strain for efficient in vivo biotinylation
CVB101- Chemically competent cells
EVB101- Electro Competent E. coli B strain for in vivo biotinylation
BIP-300 – positive control plasmid for induction
Avidity ATUM AviTag vectors and other plasmids
pBirAcm – plasmid for creation of BirA-overproducing E.coli cell line
| Vector product name(Order) | Promoter | N- or C- terminal AviTag | Short, Flexible linker(SF) or Rigid, Extended linker (RE) |
| p3A-T5-N-Avtg-SF | T5 | N- | SF (AviTag-flexible linker-your protein) |
| p3A-T5-N-Avtg-RE | T5 | N- | RE (AviTag-stiff linker-your protein) |
| p3A-T5-C-Avtg-SF | T5 | -C | SF (your protein-flexible linker-AviTag) |
| p3A-T5-C-Avtg-RE | T5 | -C | RE (your protein-stiff linker-AviTag) |
| p3A-Rha-N-Avtg-SF | Rha | N- | SF (AviTag-flexible linker-your protein) |
| p3A-Rha-N-Avtg-RE | Rha | N- | RE (Avitag-stiff linker-your protein) |
| p3A-Rha-C-Avtg-SF | Rha | -C | SF (your protein-flexible linker-AviTag) |
| p3A-Rha-C-Avtg-RE | Rha | -C | RE (your protein-stiff linker-AviTag) |
Purification and Identification of Avitag’d proteins
AbC- mab antibody against C-terminus AviTag
Purified Proteins and support chemicals
Biotinylated Twinkle� CFP – 100 ug
Biotinylated Holly� CFP – 100 ug