Biotin Agarose
$70.00 - $735.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: BIOAG-1 (for 1ml)
Cat. No.: BIOAG-5 (for 5ml)
Cat. No.: BIOAG-20 (for 20ml)
Description
Biotin Agarose, also known as Biotin Agarose Gel or Biotin Agarose Resin, is composed of high-quality biotin covalently coupled with highly cross-linked 6% agarose. This product enables rapid, efficient, sensitive, and specific binding with streptavidin or avidin, as well as antibodies, nucleic acids, proteins, peptides, and lectins that are conjugated with streptavidin or avidin. It is primarily used for the separation and purification of nucleic acids, antibodies, proteins, or related complexes that are conjugated with avidin or streptavidin, and is applicable in immunoprecipitation (IP), cell sorting, and DNA-protein interaction studies.
Biotin, also known as Vitamin B7, D-Biotin, Vitamin H, or Coenzyme R, is one of the B vitamins. Biotin is classified as a heterocyclic compound, featuring a sulfur-containing ring fused with a ureido group and a tetrahydrothiophene group. The ureido ring containing the -N-CO-N- group acts as a carrier of carbon dioxide in carboxylation reactions. Biotin plays a vital role in the metabolism of carbohydrates, fatty acid synthesis, and gluconeogenesis. The biotinylation of histones in nuclear chromatin is important for chromatin stability and gene expression. Additionally, biotin acts as a cofactor for carboxylases, such as pyruvate carboxylase, catalyzing the formation of oxaloacetate from pyruvate and carbon dioxide.
As a small molecule, biotin does not interfere with the biological functions of larger molecules. It is widely used in the biotin-(strept)avidin system and serves as an important cofactor for carboxylases, present in various metabolic pathways. The binding of biotin to avidin or streptavidin facilitates the connection of target molecules (such as antibodies, nucleotides, and protein A) with labeling systems (including enzymes, fluorescence, and chemiluminescent probes). These complexes are utilized in many detection systems, such as immunoprecipitation, immunohistochemistry, flow cytometry, and Southern and Northern blotting. Furthermore, this method is suitable for the purification and characterization of various target molecules. Biotin is also used to culture oligodendrocytes and as a vitamin supplement for the growth of Bacillus species, as well as to block endogenous biotin in immunohistochemistry processes.
Biotin agarose gel has extensive applications in the biomedicine field, allowing for the specific binding of (strept)avidin-conjugated antigens or antibodies, serving as a carrier for immunoprecipitation, cell sorting, and ELISA reactions. It can bind to (strept)avidin-conjugated DNA or RNA fragments to isolate specific nucleic acid-protein complexes from cell or tissue extracts for studies of protein-nucleic acid interactions. Additionally, it is used with (strept)avidin-conjugated nucleic acid probes for DNA and RNA hybridization experiments, or for the separation and purification of mRNA. It is also suitable for purifying single-stranded (strept)avidin-conjugated DNA oligonucleotides and isolating (strept)avidin-conjugated PCR products.
Features
- This product has a high binding capacity. Compared to many similar products, it offers an exceptionally high binding capacity, allowing for the rapid separation and purification of streptavidin-conjugated molecules in complex samples. The Biotin Agarose in this product is a 50% gel suspension, with each milliliter of Biotin Agarose containing 1-1.5 μmol of biotin, which can bind approximately 10-20 mg of streptavidin, making it highly effective for experiments such as immunoprecipitation.
- This product has strong specificity. It can specifically bind to streptavidin-conjugated ligands such as antibodies, nucleic acids, proteins, peptides, and lectins, resulting in high-purity products that can be further used in a series of subsequent analyses, including Western blotting, ELISA, Northern blotting, qPCR, and mass spectrometry.
- This product is easy to use. It is stored in a special protective solution and is glycerol-free, making it convenient to handle. The product is suitable for both small sample detection and automated high-throughput screening systems, ensuring high consistency across different operational methods.
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This product is a 50% gel suspension, with a total volume that includes 0.5 ml of gel precipitate per milliliter. If 50 μl of the agarose gel suspension is used for each sample, one milliliter of this product can be used for 20 sample reactions.
Storage
Store at -20ºC for one year of effectiveness. At 4ºC, it remains effective for at least one month.
Precautions
- This product should be adequately resuspended before use by gently inverting it several times to ensure uniform mixing of the agarose gel. Avoid vigorous vortexing or shaking, as this may cause protein denaturation or fragmentation of the agarose gel.
- The product contains trace amounts of preservatives; it is recommended to wash the gel three times with an appropriate solution such as TBS to eliminate any potential interference from the preservatives.
- When performing immunoprecipitation or purification, it is advisable to design positive and negative control groups.
- The type, size, and coupling method of the streptavidin or avidin-conjugated molecules can all affect binding efficiency. It is suggested to determine the appropriate agarose gel amount for each specific application using a gradient dilution method, and consider increasing the agarose gel amount to 2-3 times the molar quantity of the target molecules to ensure adequate binding.
- Free streptavidin may reduce the binding capacity of this agarose gel to the target protein complexes; therefore, excess free streptavidin should be removed using appropriate methods after coupling with proteins or nucleic acids.
- Once protein samples are collected, purification should be completed as soon as possible, and samples should always be kept at 4ºC or on ice to minimize protein degradation or denaturation. To effectively inhibit protein degradation, a suitable mixture of protease inhibitors can be added to the protein samples.
- If centrifugation does not completely remove insoluble materials from the protein samples, the sample solution can be filtered using a 0.45 μm filter membrane. Agarose gels that have been eluted with acidic solutions or using SDS-PAGE cannot be reused. To minimize the loss of biotin, the low pH elution step should not exceed 10 minutes, whether performed manually or automatically.
- High concentrations of DTT, β-mercaptoethanol, guanidine hydrochloride, etc., may affect the binding of this product to the ligands.
- This product is intended for scientific research use by professionals only and should not be used for clinical diagnosis or treatment, nor for food or pharmaceuticals, and should not be stored in residential areas.
- For your safety and health, please wear a lab coat and disposable gloves during handling.
Only for research and not intended for treatment of humans or animals
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