Streptavidin, His
Streptavidin Recombinant, His Tag
Cat. No.
BT3997
Source
Escherichia Coli.
Synonyms
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 95.0% as determined by analysis by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description
Recombinant Streptomyces Avidinii Streptavidin produced in E.Coli is a single, non-glycosylated polypeptide chain (25-183) containing a total of 167 amino acids and having a molecular mass of 17kDa. The Streptavidin protein is fused to an 8 aa N-terminal His-Tag and purified by proprietary chromatographic techniques.
Product Specs
Introduction
Streptavidin, a tetrameric protein derived from Streptomyces avidinii, exhibits strong binding affinity for biotin. This characteristic makes it a valuable tool in molecular biology. With a dissociation constant (Kd) of approximately 10^-15 mol/L, the biotin-streptavidin complex is highly stable. This robust interaction has led to the widespread use of streptavidin in diagnostic and laboratory kits. The streptavidin/biotin system boasts one of the largest known free energies of association for noncovalent binding between a protein and small ligand in aqueous solution (K_assoc = 10^14), highlighting its exceptional stability across various temperatures and pH levels.
Description
This recombinant Streptomyces Avidinii Streptavidin is produced in E. coli. It exists as a single, non-glycosylated polypeptide chain with a sequence spanning from amino acid 25 to 183. This 167-amino acid protein has a molecular weight of 17 kDa. An 8-amino acid His-Tag is fused to the N-terminus of the protein to facilitate purification, which is achieved through proprietary chromatographic methods.
Physical Appearance
Clear, colorless solution that has been sterilized by filtration.
Formulation
The Streptavidin protein is supplied in a solution at a concentration of 1 mg/ml, buffered in 20mM Tris-HCl at a pH of 7.5.
Stability
For optimal storage, keep the streptavidin at 4°C if it will be used within 2-4 weeks. For longer-term storage, freeze the solution at -20°C. Adding a carrier protein like HSA or BSA (0.1%) is recommended for extended storage. Repeated freezing and thawing should be minimized.
Purity
The purity of this Streptavidin is greater than 95%, as determined by SDS-PAGE analysis.
Source
Escherichia Coli.
Amino Acid Sequence
MVHHHHHHDP SKDSKAQVSA AEAGITGTWY NQLGSTFIVT AGADGALTGT YESAVGNAES RYVLTGRYDS APATDGSGTA LGWTVAWKNN YRNAHSATTW SGQYVGGAEA RINTQWLLTS GTTEANAWKS TLVGHDTFTK VKPSAASIDA AKKAGVNNGN PLDAVQQ.
Product Science Overview
Recombinant Streptavidin
Recombinant streptavidin is produced using Escherichia coli (E. coli) as a host. This recombinant form is typically engineered to be a shortened variant, often referred to as core streptavidin, which includes amino acids 13-139 . This variant is non-glycosylated and has a molecular mass of approximately 14.4 kDa per monomer, resulting in a tetrameric molecular weight of around 57.7 kDa .
His Tag
The His Tag (Histidine Tag) is a sequence of histidine residues (usually six) added to proteins to facilitate their purification. The His Tag binds strongly to nickel or cobalt ions, allowing the tagged protein to be isolated from a mixture using immobilized metal affinity chromatography (IMAC). In the case of recombinant streptavidin, the His Tag is typically added to the N-terminus of the protein .
Applications
Streptavidin recombinant with a His Tag is used in various applications, including:
- Affinity Protein Purification: The His Tag allows for easy purification of the protein using IMAC, while the streptavidin component can bind biotinylated molecules with high affinity.
- Immunoassays: Streptavidin-biotin interactions are used in various immunoassays to detect the presence of specific proteins or other molecules.
- Histochemistry and Fluorescence In Situ Hybridization (FISH): Streptavidin can be used to detect biotinylated probes in tissue samples.
- Flow Cytometry and Microarrays: Streptavidin is used to capture biotinylated molecules in flow cytometry and microarray experiments.
- Blot Analysis: Streptavidin can be used to detect biotinylated proteins on blots .
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