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Chemical Name: 6-[6-(Biotinylamino)hexanoylamino]hexanoic acid N-hydroxysulfosuccinimide ester
CAS: 180028-78-8(free acid)
Appearance: White or pale reddish-brown powder
Purity: ≥95.0% (HPLC)
MW: 669.75, C26H40N5NaO10S2
Storage Condition: -20oC
Shipping Condition: blue ice
Product Description of Amine-Reactive Biotins
avidin-biotin system has many applications in immunology and
histochemistry. The interaction between avidin and biotin is remarkably
strong with a dissociation constant on the order of 10-15
M. Biotin is usually added to primary or secondary antibodies such as
anti-IgG and anti-IgM. After preparing the antigen-antibody complex with
the biotin-labeled antibody, colorimetric, or fluorometric detection of
the antigen is performed using enzyme or fluorescein-labeled avidin or
streptavidin. Succinimidyl ester biotins react with primary and
secondary amines, such as amino acids and proteins, at pH 7-9.
Succinimidyl ester reacts with free amine groups to create a stable
amide bond. Succinimidyl biotin
reagents must be dissolved in DMSO, DMF, or alcohol. Stock solutions
prepared with DMSO are stable for several months at -20ºC. Sulfo
succinimidyl biotin reagents are soluble in water, so there is no need
to use organic solvents such as DMF or DMSO. IgG prepared using biotin
with a longer spacer such as Biotin-(AC5)2-OSu or Biotin-(AC5)2-Sulfo-OSu, has a better signal-to-noise ratio. The longer spacer enables streptavidin or anti-biotin IgG to recognize biotin without structural inhibition. Therefore, Biotin-(AC5)2-OSu is utilized as the biotin labeling agent in the Biotin Labeling Kit-NH2.
Labeling Procedure for IgG
1. Prepare 10 mM of the biotin labeling reagent using DMSO.
2. Prepare 100 μl of 1 mg per ml IgG buffer solution (pH 7.5-8.5) that does not contain any large molecules with amine compounds.
3. Add 1-5ml biotin labeling reagent DMSO solution to the IgG buffer solution and incubate at 37ºC for 1 hour.
4. Remove excess biotin labeling reagent using gel filtration or dialysis.
5. Prepare solutions for further experiment using an appropriate buffer such as PBST (0.05% Tween 20/PBS).
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Nucleotide: Usefulness in the Recovery of Protein-DNA Complexes from
Avidin Affinity Columns, Proc. Natl. Acad. Sci. USA, 1985, 82, 2593.
W. J. LaRochelle and S. C. Froehner, Immunochemical Detection of
Proteins Biotinylated on Nitrocellulose Replicas, J. Immunol. Methods,
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All Products are applicable only for life science research. Not for diagnostic research use.
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