LPAP (Lymphocyte Phosphatase-Associated Phosphoprotein) Introduction

Human LPAP（Lymphocyte Phosphatase-Associated Phosphoprotein） is a phosphoprotein of alien action that noncovalently assembly with CD45 in lymphocytes. In CD45-deficient animal T cells, LPAP protein is actinic at accustomed levels but is added rapidly base than in wild-type cells. Announcement of CD45 cDNA rescues LPAP protein expression. This acerb suggests that LPAP is adequate from abasement through its alternation with CD45. We accept mapped the sites of alternation amid LPAP and CD45 employing chimeric CD45 molecules and LPAP abatement mutants. Our abstracts authenticate that the alternation amid LPAP and CD45 is advised via the transmembrane regions of both molecules. In addition, the intracytoplasmic amino acids adjoining to the transmembrane arena of LPAP may access its bounden to CD45.

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Insulin discovery process

Insulin is a pancreatic β cells by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, such as glucagon stimulated secretion of a protein hormone. Insulin is the only way to reduce blood glucose in the body hormones, while promoting glycogen, fat, protein synthesis. Exogenous insulin is mainly used to treat diabetes, diabetic patients with early use of insulin and powerful antioxidants such as (alpha lipoic acid for injection, oral astaxanthin, etc.) is expected to appear longer honeymoon period, there will be no addiction and insulin injections dependence.
Discovery process

Insulin in 1921 by the Canadian FG Banting and CH Best first discovered. 1922 began to be used clinically, so that diabetic patients died in the past have been saved. Indications Kidney Institute, Chinese Academy of detection until the early 1980s, almost all insulin for clinical from pigs, cattle pancreas extract. Insulin composition of different animals in both differences pig and human insulin is most similar to the structure, only the B-chain of a different amino acids carboxyl terminal. In the early 1980s has been successfully used genetic engineering techniques of mass production by microorganisms human insulin, and has been used clinically.
1955 British F. Sanger team determined the complete amino acid sequence of bovine insulin, human knowledge has opened up the road of the chemical structure of protein molecules. September 17, 1965, Chinese scientists have synthesized a full biological activity of crystalline bovine insulin, it is the first one in the laboratory using artificial methods of protein synthesis and later the United States and the Federal German scientists also completed a similar work.
The early 1970s, the UK and Chinese scientists has successfully used X-ray diffraction method of three-dimensional structure of porcine insulin. These efforts for the further study of molecular structure and function of insulin relationship foundation. People use chemical synthesis and semi-synthetic analogs prepared to study the structural changes on the biological functions of; for different species of comparative study of insulin; study disease abnormal insulin molecule, i.e. the insulin gene mutations of individual amino acids in the insulin molecule arising from a change in molecular disease. These studies illustrate some of the causes of diabetes also has important practical significance.



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Macrophage culture method

Macrophage is located within tissue leukocytes, monocytes derived cells, and monocytes from the bone marrow and the precursor cells. Macrophages and monocytes are phagocytic cells, non-specific defense in vertebrates (innate) and specific defense (immune). Their main function is to fix the cells in the form of free cells or cell debris and the phagocytosis of pathogens (i.e. phagocytosis and digestion), and activated lymphocytes or other immune cells, making it react to pathogens.
Culture methods
Macrophage cell line also has unlimited, mostly from the mouse, such as P338D1, S774A.1, RAW309Cr.l, etc., were vicious, macrophages were cultured in cell morphology and phagocytic function, easy passage and sidewall separation, but difficult to construct strains. Cultured macrophages in various ways and from various sources available to get the cells to mice derived method is most useful, and its method is as follows:
1, three days before the experiment, each mouse was intraperitoneally injected to thiolacetic acid sterile broth 1ml (Do not inject the intestines).
2, hoping to kill animals, portable mouse tail will all be immersed in 70% ethanol for 3 to 5 seconds.
3, set the animal on the autopsy table, with a needle fixed limbs, hands holding tweezers pull to the side torn skin, exposing the peritoneum, but not to hurt the peritoneal wall.
4, and then 70% alcohol scrub peritoneal wall, the use of a syringe was injected into the abdominal cavity suction 10mlEagle while rubbing pressure from both sides of the peritoneal wall with your fingers, so that adequate flow of fluid in the abdominal cavity.
5, gently stir the abdominal wall with a needle, the animal body slightly to one side, so that the fluid collection in the abdominal cavity under the needle into the needle tube drawing.
6, carefully pull out the needle, inject the liquid centrifuge tube, 250g4 ℃ for 10 min, the supernatant, add 10mlEagle medium.
7, cells were counted per mouse can produce 20 ~ 30 × 105 cells, of which 90% macrophages.
8, to obtain tie with 3 × 105 cells / square centimeter, to be inoculated with 2.5 × 106/ml. 9, the cell culture was purified to remove other white blood cells, a few hours after inoculation, culture solution, rinsed with Eagle 1 to 2 times, the addition of a new culture medium Eagle set 37 ℃ CO2 culture incubator.

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Lysozyme preparation

Lysozyme, also known as muramidase (muramidase) or N-acetyl murein polysaccharide hydrolase (N-acetylmuramide glycanohydrlase), is a kind of bacteria in the hydrolysis of the enzyme alkaline mucopolysaccharides. Primarily through the destruction of the cell wall of N-acetyl muramic acid and N-acetyl glucosamine between the β-1, 4 glycosidic bond, so that the insoluble cell wall mucopolysaccharides into soluble glycopeptides contents escape rupture cell walls leaving bacteria dissolved. Lysozyme can be negatively charged with direct binding of viral proteins, and DNA, RNA, apoprotein form complex salts, the virus inactivation. Thus, the enzyme has antibacterial, anti-inflammatory, anti-viral effects.
Lysozyme is a clone of biological engineering technology, extraction and preparation, it is a natural enzyme, safety green additive, no resistance.
The enzyme exists in a variety of human tissues, birds and poultry egg white, mammal tears, saliva, plasma, urine, milk and other body fluids and micro-organisms also contain this enzyme, of which the most abundant egg white. Extracted from the isolated egg lysozyme is 18 kinds of amino acid residues 129 a single polypeptide chain. It is rich in basic amino acids, 4 bonds maintain the enzyme conformation, is a basic protein, the N-terminal lysine, C terminal leucine. Decomposable Micrococcus lysodeikticus, Bacillus megaterium, yellow Sarcina other Gram-positive bacteria.
Lysozyme less than the isoelectric point wide pH range, with a positive charge molecules can be adsorbed on the weakly acidic cation exchange resin, eluted salting, lysozyme can be obtained precipitate was then purified product can be obtained. Adsorption resin egg wash ↓ 724 ↓ ↓ eluted with a buffer solution of ammonium sulfate salting-dialysis ↓ NaOH to basic protein precipitate freeze dried powder was dried lysozyme Lysozyme 5 ~ 10 ℃ under 540kg fresh egg white added to the processed Good 80kg 724 resin and stirred adsorbed 6h, allowed to stand at 0 ~ 5 ℃ overnight. Decanted upper egg white, centrifugal drying resin, washed repeatedly with distilled water adhesion egg protein, and the resin into the column with a 0? 15mol / L, pH = 6? 5 phosphate buffer solution, about 150L resin was washed, then Approximately 10% ammonium sulfate solution 600L eluate. Eluate was added ammonium sulfate to give a final volume of 40% with ammonium sulfate, a white precipitate formed, cool place overnight. Siphon supernatant drained precipitate filtered off with suction, dissolved in distilled water and then with 1 X precipitate was thin paste, and then into the dialysis bags, about 5 ℃ conditions, dialyzed against distilled water for about 24h, the intermediate changing the water 2-3 times . Precipitate was removed by centrifugation, and then a small amount of precipitate was washed once by centrifugation was combined with the washings. Beyond dialysis supernatant was slowly added 1mol / L sodium hydroxide solution, while constantly stirring, the pH increased to 8? 0 ~ 9? 0, if the white precipitate is removed by centrifugation. Then 3mol / L hydrochloric acid to adjust pH = 5? 0, freeze-drying, white sheet of lysozyme. Can also be used centrifugate 3mol / L hydrochloric acid pH = 3? 5, was added slowly with stirring a 5% solid sodium chloride, at a temperature of about 5 ℃ stand 48h, centrifuged, washed with acetone precipitate 0 ℃, drying, lysozyme.


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For Prostate Stem Cell Antigen-related information

Prostate stem cell antigen (PSCA) is a 1998 Reiter et al study gene expression in prostate cancer discovered during a tumor-associated antigen, belonging Ly-6/Thy-1 family, is a 123 amino acid glycoprotein. Because PSCA with stem cell antigen -2 (SCA-2) having a homology of 30%, which is called the prostate "stem cell antigen", but the current study, PSCA expressed mainly in cell differentiation and maturation of stem cells than the surface of the surface, their specific function is unclear. PSCA in prostate cancer, bladder cancer, and pancreatic cancer, increased expression and found its expression levels and prostate cancer clinical stage and metastasis, so PSCA has become of these tumor diagnosis and prognosis of biological markers. In addition to PSCA also become the target of these tumors immunotherapy Points, particularly in the treatment of prostate cancer, immune show great potential for clinical application. PSCA as a target in prostate cancer immunotherapy preclinical studies, the use of transgenic mice or mouse prostate cancer model for preclinical evaluation. To further explore the purpose of PSCA in prostate cancer immune RT-PCR amplification of mouse prostate stem cell antigen (mPSCA) gene prokaryotic expression vector pET-42a-mPSCA, induced mPSCA protein expression and purification, and to detect mPSCA antigenic activity. Methods RT-PCR method from the mouse prostate cancer cell line RM-1 cells was amplified mPSCA gene sequencing PCR method to remove mPSCA after the signal peptide sequence was cloned into the prokaryotic expression vector pET-42a to construct recombinant plasmid pET -42a-mPSCA. To convert To E. coli BL21 (DE3) induced expression, the fusion protein is then purified. Analyzed by SDS-PAGE, Western blot detection of purified protein, the purified protein further package board after its antigenic activity by ELISA were evaluated. Results sequencing results confirmed length mPSCA successfully amplified gene; digestion and sequencing Results confirmed pET-42a-mPSCA prokaryotic expression vector was successfully constructed; transformation can be successfully induced and purified proteins consistent with the expected size; Western blot test confirmed the purified protein could react with the specific antibody; ELISA assay showed purified mPSCA immunogenic antigens. Conclusion successfully amplified full-length mouse PSCA gene was successfully constructed mPSCA prokaryotic expression vector obtained purified mPSCA protein, which has a good antigenicity for further research to PSCA as a target for immunotherapy of prostate cancer foundation.


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Anti-FBN1 (fibrillin 1) basic information

Product name: Anti-FBN1 (fibrillin 1)
Description: Fibrillin-1 is a protein that in humans is encoded by the FBN1 gene.
This gene encodes a member of the fibrillin family. The encoded protein is a large, extracellular matrix glycoprotein that serve as a structural component of 10-12 nm calcium-binding microfibrils. These microfibrils provide force bearing structural support in elastic and nonelastic connective tissue throughout the body. Mutations in this gene are associated with Marfan syndrome, isolated ectopia lentis, autosomal dominant Weill-Marchesani syndrome, MASS syndrome, and Shprintzen-Goldberg craniosynostosis syndrome.


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Anti-HEV in some applications

Anti-HEV IgM and IgG can be divided into two kinds of antibodies, Anti-HEV IgM and recent infection, infection in the early liver E (around the beginning of the acute phase) appears to last about five months and then disappears. Anti-HEV IgG appeared later than IgM, may be present in the body at least 5 years of infection indicators for ever. But many studies have shown, E liver antibody (either IgM or IgG) is not a good indicator of infection, since confirmed by PCR in patients with acute liver E, often undetectable Anti-HEV IgM or IgG, expressed antibody sensitivity is poor. Therefore, only the reference antibody measurement results can not be used as a basis for diagnosis.
E liver virus and hepatitis A infection pathway similar to fecal-oral infection based. E liver infection mostly sporadic cases of outbreak (outbreaks) occur rarely. Study found that regional epidemic E liver and contaminated water mostly related to improvement in water control, the new cases disappear. E liver incubation period of about 2 to 10 weeks, the majority of patients with flu-like symptoms appear, a few evolve into acute hepatitis, but it will not develop into chronic hepatitis or cirrhosis.
E and A liver infection liver age is slightly different, A liver infection in childhood when, and E in the liver and more infected between the ages of 15-40. Hepatitis E cases, cases of acute deterioration of liver more than A, especially pregnant women infected with hepatitis E, its mortality rate can be as high as 20%.


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Anti-HCV-Core of some description

HCV gene structure:
HCV is a positive spherical enveloped single-stranded RNA viruses. HCV genome is a single strand RNA, total length of about 9500 bases. Genome sides were 5 'and 3' non-coding region, in the middle of an open reading frame (ORF), divided into structural and non-structural region area (Figure 1). Structural protein region including the core region (C) and two envelope protein region (E1, E2), respectively, encoding core protein and envelope proteins. Non-structural protein region including NS2, NS3, NS4 and NS5 region encoding functional proteins, such as proteases (NS2, NS3 and NS4A area), helicase (NS3) and RNA-dependent RNA polymerase (NS5B region). Although the non-structural protein is not part of the virus particle, but in viral replication plays a very important role. Now commonly used genetically engineered to express the structural and non-structural proteins as coating antigen to create anti-HCV ELISA detection method. HCV genome has significant heterogeneity, different sections of the same degree of variation in the genome there are significant differences. 5 'non-coding region most conservative, so PCR detection of HCV, RNA primer design more choice in the region.

Anti-HCV (Anti-HCV-Core) is the hepatitis C virus antibodies.
It is currently the most widely used for epidemiological investigation, clinical hepatitis C screening and diagnostic test items. So far the method detection of anti-HCV continuous improvement. With the improved method, the sensitivity and specificity of a gradual increase.
Referred to as the hepatitis C virus HCV, the classification of the virus are positive-strand RNA viruses, is considered to be caused by the early non-A, non-B hepatitis virus (non-A, non-B viral hepatitis) one of the major virulence factors. Was discovered in 1978, in 1989 obtained by gene technology was first genome sequence. It is a large 45 nm, with a housing, the single-stranded RNA virus, which belongs to the Flaviviridae virus is hepatitis. According to the 1994 establishment of the new classification system, HCV can be divided into six different genotypes (Genotype) and 30 subtypes (subtype). Mainly in Europe and Japan, mainly type 1, and the second type in Japan, China, mostly in the third type in Thailand, Singapore, India, and most of the more common in Egypt, the Middle East and central Africa, mainly type 4, The Type 5 and Type 6 is more common in South Africa and Southeast Asia. Hepatitis C only infect humans. As the name implies, namely anti-HCV hepatitis C virus antibodies.

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Anti-HO-2 (Heme oxygenase 2) information about the

Heme oxygenase (Anti-HO-2 (Heme oxygenase 2)) is in the process of heme catabolism rate-limiting enzyme in the human body of CO is dominated by heme oxygenase (HO) metabolites produced. HO There are three types: oxidative stress inducible (HO-1), constitutive (HO-2) and remained clear HO-3. Studies show that, HO-1 in the body not only play a role in the physiological state, but mainly in the body, or other non-normal stress play a role. HO-2 is mainly distributed in the central nervous system and testicular, HO-2 CO produced in the nerve plays an important role in signal transduction, and CO neurotransmitters play closely related. Many scholars on ways to produce CO and HO carried out a detailed study that through this - pathways of CO speed and quantity of 0.4ml / h (16.4/μmol/h). It is known HO distributed in almost all the organs and tissues, and involves the growth process, which prompts the production of CO can be very wide area. Currently a large number of studies have confirmed CO is important in vivo cell messenger, heart, brain and vascular system plays an important biological effect, involved in the regulation of many physiological and pathological processes in vivo. Meanwhile, CO as a novel neurotransmitter is also cause for concern, and its role in the nervous system and its clinical significance in research, neuroscience has become a new hotspot.

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mGluR5 (Anti-GRM5 (Glutamate Receptor Metabotropic 5)) gene structure

First reported in 1985 Glu Sladeczek activation of G protein-coupled receptors. 1991 Houamed etc. from rat cerebellum cDNA library cloned out of the mGluR1. 1992 年 researchers mGluR1 probe made ​​from mouse brain cDNA library isolated mGluR5 (Anti-GRM5 (Glutamate Receptor Metabotropic 5)）. mGluR5 length of the cDNA is 3 918bp, encoding a 171 amino acid residues, a molecular weight of 128 289. mGluR5 consists of three parts: The first part is the function of the extracellular region, including the 560 amino acid residues, which has the potential glycosylation of tion sites, a signal peptide of the N-terminal; second portion by a hydrophobic region 7 formed in the cell membrane seven transmembrane region, this area has 261 amino acid residues; third part is the function of the cell area, the 350 amino acid residues, the C-terminal phosphorylation sites are many and threonine, serine residue, suggests that this region may be adjusted several kinase receptor target [3] in the human and rat mGluR5 7th a transmembrane region of the cytoplasm and after the insertion end 50, respectively, and 32 amino acid residues, does not cause GluR5 pharmacological properties of the changes, which will initially be cloned mGluR5 named mGluR5a, the named after insertion mG1uR5 mGluR5b, in the adult rat brain mG1uR5b mRNA expression than mGluR5a.

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