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BTAG

Sigma-Aldrich

BiotinTag™ Micro Biotinylation Kit

Synonyme(s) :

Biotinylation Kits

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About This Item

Code UNSPSC :
12352200
Nomenclature NACRES :
NA.32

Technique(s)

bioconjugation: suitable

Température de stockage

2-8°C

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Description générale

For custom biotinylation of proteins, Sigma offers kits for conjugation on two different scales. The biotin-avidin system has become popular when high sensitivity and specificity are desired. Biotin can be conjugated to antibodies, lectins, enzymes, and other proteins. The protocols in the kits have been optimized for antibodies. Avidin binds to biotin with a high affinity (Ka = 1015 M) and specificity. When conjugated to enzymes or fluorochromes avidin provides a means of identifying biotinylated compounds via enzymatic conversion of substrate to form a visible product or detection of fluorescence by spectrophotometry or flow cytometry.

Application

BiotinTag™ Micro Biotinylation Kit has been used to generate biotylanted IgG2a antibody conjugate and in the biotinylation of interleukin 9 receptor α (IL-9Rα) and thymic stromal lymphopoietin α (TSLPRα).
Biotin has been modified with aminocaproate, then activated via an ester linkage with sulfo-N-hydroxysuccinimide (BAC-Sulfo-NHS). Aminocaproate provides a six-carbon spacer that reduces steric hindrance on the biotin and improves accessibility to the binding site on avidin. Sulfonation of the hydroxysuccinimide increases the polarity of the reagent, allowing it to dissolve easily in aqueous buffer. The ester provides a carbonyl carbon adjacent to a labile ester linkage as a target for primary amine side chains of accessible lysine residues, joining the biotinamidocaproate to the protein via an amide bond.

Caractéristiques et avantages

  • Complete protocols for labeling and assay
  • BAC-Sulfo-NHS is completely water soluble
  • No DMF or DMSO needed
  • Biotinylation occurs near neutral pH and physiological ionic strength, avoiding harsh conditions that could damage sensitive proteins
  • Fast separation of conjugate from reactants using gel filtration
  • 2-5 molar ratio biotin to protein in conjugate (for antibodies)
• Two scales to choose from:
1 mg protein per reaction (B-TAG)
10 mg protein per reaction (BK-101)
• Sufficient reagents for at least 5 labelings

Remarque sur l'analyse

Procedure:
Conjugation is performed in four easy steps:
1. Reconstitute BAC-Sulfo-NHS with Phosphate Buffer (PB).
2. Add BAC-Sulfo-NHS to protein and allow to react for 30 minutes at room temperature.
3. Separate the conjugate from the reactants on gel filtration.
4. Assay the conjugate for biotin incorporation by the avidin-HABA assay (BK-101 only). Conjugate is ready to use.

Informations légales

BiotinTag is a trademark of Sigma-Aldrich Co. LLC

Composants de kit également disponibles séparément

Réf. du produit
Description
FDS
  • P38130.01 M PBS, pH 7.4, powder 1 LFDS
  • P38130.1 M phosphate buffer, pH 7.2, powder 1 x 5FDS

Pictogrammes

Exclamation mark
GHS07

Mention d'avertissement

Warning

Mentions de danger

H317

Classification des risques

Skin Sens. 1

Code de la classe de stockage

12 - Non Combustible Liquids

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Consulter la Bibliothèque de documents

Quantum dot enabled lateral flow immunoassay for detection of cardiac biomarker NT-proBNP
Wilkins MD, et al.
Sensing and Bio-Sensing Research, 21, 46-53 (2018)
Carole L Browne et al.
The Journal of experimental biology, 210(Pt 7), 1275-1287 (2007-03-21)
Numerous reports document that the 70 kDa heat shock proteins are not only intracellular proteins but are also present in blood and other extracellular compartments. How they affect cell function from the extracellular space remains unclear. Using two well-characterized cell
Ming Yang et al.
Viruses, 14(4) (2022-04-24)
Foot-and-mouth disease virus (FMDV) causes FMD, a highly contagious disease of cloven-hoofed animals including cattle, goats, pigs and sheep. Rapid detection of FMDV is critical to limit the devastating economic losses due to FMD. Current laboratory methods for FMDV detection
IL-2,-7, and-15, but not thymic stromal lymphopoeitin, redundantly govern CD4+ Foxp3+ regulatory T cell development
Vang KB, et al.
Journal of Immunology, 181(5), 3285-3290 (2008)

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