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How many kinds of common biochemical reagents are there?
2019-03-04
来源：亚科官网
With the development of life sciences, biochemical reagents have developed into a large class of chemical reagents, with more than 10,000 kinds of commodities. There are more than 2,500 biochemical reagents sold in China. At present, biochemical reagents cover liver function, kidney function, blood lipids, heart muscle, pancreas, special proteins, etc., which can meet the various biochemical detection needs of end users.
The concept of biochemical reagent
A biochemical reagent is an essential component of an organism extracted from a living body or chemically synthesized, and is used for analysis and identification of biological components and manufacture of biological products. Biochemical reagents are mainly diagnostic reagents for blood routine, urine routine, liver function, kidney function, pancreas, diabetes and other diseases. Biochemical diagnosis is the first automated detection method and one of the most commonly used in vitro diagnostic methods.
Common biochemical reagent types
1. Proteinase K
It can hydrolyze and digest proteins, especially histones that bind to DNA. It is stable in urea and SDS. The typical working concentration is 50~100 μg/mL. The recommended reaction buffer is 50mM Tris-HCl (pH 7.5), 10 mM CaCl2 .
2. SDS
Sodium lauryl sulfate dissolves lipids and proteins on the cell membrane, thereby dissolving membrane proteins and destroying the cell membrane, and dissociating the nuclear proteins in the cells. SDS can also bind to proteins and precipitate.
3. IPTG
Isopropyl-β-D-thiogalactoside, commonly used for blue-white spot screening and IPTG-induced protein expression in bacteria, etc. IPTG and lactose are structurally similar, so it can be repressed with lactose operon, the spatial conformation of the repressor protein changes, the tetramer depolymerizes into a monomer, loses the ability to specifically bind to the operon, thereby releasing the repressor protein, allowing the subsequent gene to be transcribed and synthesizing the enzyme using lactose.
Unlike lactose, IPTG is not hydrolyzed by β-galactosidase. It often used with X-GAL for blue and white spot screening. An extremely powerful inducer that is not metabolized by bacteria and is very stable.
4. MOPS
A buffer that inhibits RNase, generally used for RNA denaturing electrophoresis together with formaldehyde (for variant RNA).
5. guanidine hydrochloride
Guanidine hydrochloride can break hydrogen bonds in the protein structure, which increases the solubility of non-polar molecules including amino acid side chains and reduces hydrophobic interactions.
6. Acridine orange
It is a kind of fluorescent pigment. It has different binding amount to DNA and RNA in cells. It can emit fluorescence of different colors. It has less green fluorescence when combined with DNA, and more orange or orange-red fluorescence when combined with RNA. The dye has membrane permeability and can permeate the cell membrane to stain nuclear DNA and RNA.
Under the fluorescence microscope, acridine orange can pass through the normal cell membrane, making the nucleus green or yellow-green uniform fluorescence; in apoptotic cells, due to chromatin condensation or rupture into fragments of different sizes, the formation of small apoptosis body. Acridine orange is stained with dense yellow-green fluorescence or yellow-green debris particles; while necrotic cells yellow fluorescence is weakened or even disappeared.
Acridine orange AO is often double-dyed with ethidium bromide EB. Because EB only kills cells to produce orange fluorescence, it can distinguish normal cells, apoptotic cells and necrotic cells. It can also be used as a mutagen for frameshift mutations, which can be inserted between two adjacent base pairs, which will increase or delete one base of the DNA strand during DNA replication, resulting in a frameshift mutation. Acridine orange dye is toxic, wear gloves when handling, and be protected from light.
7. DMSO
The dimethyl sulfoxide is commonly used as a solvent for liquid chromatography, and is used as a reference material for the UV extinction value of the test substance, and is soluble in all alkanes and olefins.
8. TRICINE
N-[tris(hydroxymethyl)methyl]glycine, the common electrophoresis system is the Tris-glycine system; Tris-Tricine electrophoresis is used for the electrophoresis of peptides and proteins with a molecular weight of less than 10kDa, which can obtain better separation effect.
Related links: MOPS
Edited by Suzhou Yacoo Science Co., Ltd.
