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In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
NHS-Biotin (C14H18O5N3S) is N-hydroxysuccinimido biotin. It is the shortest of three similar EZ-Link NHS-Biotin Reagents that enable simple and efficient biotinylation of antibodies, proteins and any other primary amine-containing biomolecules in solution. NHS-Biotin offers researchers the possibility of optimizing labeling and detection experiments where steric hindrance of biotin binding is an important factor. Because it is uncharged and contain simple alkyl-chain spacer arms, NHS-Biotin compound is membrane-permeable and useful for intracellular labeling.
NHS-Biotin is the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.
Ref:
• Protein labeling—biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins • Intracellular labeling—membrane-permeable, can be used to label inside cells • Amine-reactive—reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the N-terminal-amine • Irreversible—forms permanent amide bonds; spacer arm cannot be cleaved • Solubility—water insoluble, must be dissolved in DMSO or DMF before further dilution in aqueous buffers • Very short—spacer arm is 13.5 angstroms; it consists of the native biotin valeric acid group only
Sample
whole blood
Preparation method
Soluble in DMSO or DMF.
Reaction Conditions
30 minutes
Applications
Firstly, NHS-biotin was dissolved in DMSO at a concentration of 100 mg/mL, after that using 90% DMSO solution diluted the biotin to a concentration of 20 mg/mL. The biotin solution was sterilized by filtering the solution through 0.2 μm pore nylon syringe filters and the solution was diluted with 6 volumes 0.9% saline solution. Blood was allowed to warm to room temperature prior to labeling with NHS-biotin. To achieve a biotin concentration of 0.04 pg of biotin/RBC（Red Blood Cells）, Biotin DMSO saline solution was injected into each blood bag and agitated for 30 minutes.
References:
[1]. J.E. Tomlinson, E. Taberner, R.C. Boston, S.D. Owens, and R.D. Nolen-Walston. Survival Time of Cross-Match Incompatible Red Blood Cells in Adult Horses. J Vet Intern Med 2015;29:1683–1688.
