| Description | Extinction Coeff.A276 nm = 0.41 at 1.0 mg/mLMolecular Weight8933 Da (single chain)Protein Purity>97% by SDS-PAGEGeneral DescriptionHuman C3a desArg is prepared from normal human serum after activation of C3 by human C3 convertase, followed by the removal of the C-terminal arginine by the natural Extinction Coeff.A276 nm = 0.41 at 1.0 mg/mLMolecular Weight8933 Da (single chain)Protein Purity>97% by SDS-PAGEGeneral DescriptionHuman C3a desArg is prepared from normal human serum after activation of C3 by human C3 convertase, followed by the removal of the C-terminal arginine by the natural carboxypeptidase N (Hugli, T.E. et al. (1981); Meuller-Ortiz, S.L., et al. (2009)). C3a is amember of the anaphylatoxin family of three proteins (C3a, C4a and C5a) produced by the activation of complement. The desArg form of C3a is an unglycosylated polypeptidecontaining 76 amino acids with a molecular mass of 8,933 daltons. C3a mediates many inflammatory responses including smooth muscle contraction, vasodilation, increased vascular permeability, and release of histamine from mast cells and basophils (Law, S.K.A. and Reid, K.B.M. (1995)). These activities of C3a are inactivated by removal of the Cterminal arginine and this is complete within minutes after formation in plasma. However, new activities have been identified for C3a desArg acting through the C5L2 receptor (Kalant, D. et al. (2005)). These activities were first assigned to ASP (acylation-stimulating protein), but this was later shown to be identical to C3a desArg. C3a desArg binds to C5L2 with a Kd of approximately 70 nM. In adipocytes, macrophages, fibroblasts and may other cellstriglyceride synthesis, glucose transport, and fatty acid uptake are stimulated upon C3a desArg binding in both humans and mice. C3 knockout mice lack ASP function andrecombinant C3a desArg is fully functional while C5L2 knockout mice are unresponsive. Many of the biological functions of insulin are expressed by C3a desArg and it also expresses endocrine effects on insulin secretion by pancreatic cells (Maslowska, M. et al (2005)). Physical Characteristics & StructureMolecular weight: 8,933 calculated molecular mass. Observed mass (MALDI-TOF) is 8,934 + 9 mass units.Amino acid sequence (76 amino acids): SVQLTEKRMD KVGKYPKELR KCCEDGMREN PMRFSCQRRT RFISLGEACK KVFLDCCNYI TELRRQHARA SHLGLA X-ray-derived crystal structure: Huber, R. et al. (1980) NMRderived structure: Nettesheim, D.G. et al. (1988); Murray, I. et al. (1999).FunctionSee General Description above. AssaysMany of the assays for C3a can be used for C3a desArg if 100- to 1000-fold higher concentrations are used, but in reality the desArg for is essentially inactive in such assays as the contraction of guinea pig ileum, permeation of a dye such as trypan blue from the vasculature into skin, mast cell degranulation, (measured as histamine release), platelet aggregation, IL-1 release from monocytes and the release of prostaglandins and leukotrienes from many cells and tissues. Similarly, ATP release from guinea pig platelets, serotonin relaease from guinea pig platelets, N-acetyl-beta-D-glucosamidase release from differentiated U937 cells and calcium release from differentiated U937 cells (Dodds, A.W. and Sim, R.B. (1997)) are all reduce to very low levels in the desArg form.The newly acquired functions of C3a desArg related to lipid metabolism detailed in the General Description section above may be assayed by any of a large number of assays detailed in the extensive literature on acylation-stimulating protein (ASP) (Kalant, D. et al. (2005); Maslowska, M., et al. (2005); Murray, I., etaal. (1999)). ELISA kits for the assay of C3a desArg levels in blood and other fluids are sold by many companies. A radioimmunoassay for C3a/C3a desArg is also available. These measurements are useful for detecting complement activation in vivo, but the interpretation of their meaning is complicated by the fact that clearance of the anaphylatoxins is rapid. In vivoFreshly drawn normal human serum contains approximately 17 nM C3a desArg (corresponding to activation of about 0.3 % of the total C3). Although this may represent the resting concentration in vivo it is difficult to draw or store blood without some C3 activation so a true in vivo concentration is difficult to determine. The presence of EDTA and Futhan in the collection tubes can minimize this background (Pfeifer, P.H. et al. (1999)). Full activation of all C3 in blood (1200 µg/mL) would result in ~6,600 nM C3a desArg (~60 µg/mL). Due to the sensitivity of the lipid metabolism stimulating functions of C3a desArgresponses can theoretically be initiated by activation of approximately 1/1,000 of the C3 in a local area.RegulationC3a desArg levels are regulated by three processes: formation, inactivation and clearance. The enzymes that cleave C3 and release C3a (collectively called C3/C5 convertases) do so at a rate approximately 300-times the rate that these enzymes cleave C5(Pangburn, M.K. and Müller-Eberhard, H.J. (1986); Rawal, N. and Pangburn, M.K. (2001)). C3a is “inactivated” by removal of its C-terminal arginine amino acid. The product C3a desArg (or C3a without the C-terminal arginine) is produced by the action of the plasma enzyme carboxypeptidase N (Mueller-Ortiz S.L. et al. (2009)). The inactivation is rapid and most C3a is converted to C3a desArg within minutes of its formation. “Inactivated” C3a still possesses some biological activities, but it is considered inactive for most C3a-specific functions. As described above C3a desArg does possess numerous activities of its own as the acylation-stimulaing protein (ASP) released by adipose tissue. Because of the large number of cells bearing C3a and C3a desArg receptors (endothelial, immune, adipose, smooth muscle, neuronal, etc.) the capture, internalization and digestion of C3a and C3a desArg results in its rapid removal from circulation.DeficienciesA deficiency of C3 or a deficiency of the enzymes that cleave C3 to generate C3a result in the absence of C3a desArg. There are no known complete deficiencies of all of the C3 convertases. Examples of C3 deficient humans (Ghannam A, et al. (2008)) and mice (Wessels, M.R. et al. (1995)) exist, but the degree to which pathologies associated with C3 deficiency are due to the lack of C3 or the absence of C3a is unclear. Information on this has been acquired from C3aR and C5L2 receptor knock-out animals (Singer, L. et al. (1994);MacLaren, R. et al. (2008); Maslowska, M. et al. (2005)). DiseasesSee sections above for many biological effects of C3a and C3a desArg connected with inflammatory reactions in many diseases and lipid metabolism. In addition, a role for C3a in asthma has been well documented (Zhang, X. and Kohl, J. (2010); Wills-Karp, M. (2007)). Diseases related to C3a desArg regulation of the triglyceride synthesis, glucose transport, insulin regulation and fatty acid uptake have not yet been described.Precautions/Toxicity/HazardsThis protein is purified from human serum and therefore precautions appropriate for handling any blood-derived product must be used even though the source was shown by certified tests to be negative for HBsAg, HTLV-I/II, STS, and for antibodies to HCV, HIV-1 and HIV-II.Hazard Code: B WGK Germany 3... Read More | The Leuconostoc GPDH exhibits dual coenzyme specificity, namely NAD and NADP (Olive and Levy, Biochem., 6, 730 730, 1967). When assayed under conditions that are optimal for the particular coenzyme, the ratio of observed catalytic activity is NAD/NADP = 1.8 | Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote the shift of reaction equilibrium to the product generation end, and increase the amount of products.The molecular weight of PPase (pyrophosphatase, inorganic, inorganic pyrophosphatase) is about 63kd, which can catalyze the hydrolysis of inorganic pyrophosphate to produce orthophosphate: P2O74_+H2O+PPase→2HPO42_. In the nucleic acid amplification experiment, PPase can hydrolyze the inorganic pyrophosphate generated with the reaction to avoid its inhibition on the reaction system. The removal of pyrophosphate can shift the reaction equilibrium to the product generation end.This product is a GMP level recombinant inorganic pyrophosphatase (yeast source) expressed by large-scale fermentation of E. coli. It is produced with raw and auxiliary materials of medicinal specifications, and the host protein residue and nucleic acid residue are strictly controlled. The product production and quality management procedures in line with GMP specifications ensure that the production process and all raw and auxiliary materials can be traced.Quality requirements project standard appearance Clear liquid Visible foreign matter Compliance with regulations PH value 7.5±8.5 activity 98U/ml-102U/ml purity ≥95% Endonuclease residues Degradation of substrate shall not exceed 10% Exonuclease residues Degradation of substrate shall not exceed 10% RNase residue Degradation of substrate shall not exceed 10% Bacterial endotoxin content ≤10EU/ml Exogenous DNA residue ≤100pg/mg Host protein residue ≤50ppm Mycoplasma detection negative Heavy metal residues ≤10ppm Follow the following specifications1. ISO 9001:2015, certified facility。2. GMP appendix - cell therapy products State Drug Administration.3. general introduction to human gene therapy - Chinese Pharmacopoeia 2020, National Pharmacopoeia Committee.4. USP chapter <1043>, adjuvant materials for cell, gene, and tissue engineered products.5. USP chapter <92>, growth factors and cytokines used in cell therapy manufacturing.6. Ph. Eur. General chapter 5.2.12, raw materials of biological origin for the production of cell-based and gene therapy medical products.Product features1. hydrolyze inorganic pyrophosphate.2. DNA synthesis: significantly enhance DNA replication ability.3. RNA synthesis: increase RNA production in in vitro transcription reaction.4. The optimal reaction temperature is 25℃, and the enzyme can be inactivated at 65℃ for 10min.Product usage1. optimize RNA transcription: improve the RNA yield of in vitro transcription reaction.2. remove PPI contamination from reagents for SNP genotyping by pyrophosphate assay.3. promote the synthesis of protein, RNA and DNA.4. catalyze the reaction of PPI + H2O → 2pi.5. ssr-pcr optimization：Improve efficiency and increase DNA production.Activity definitionCatalytic inorganic pyrophosphate formation 1 per minute under standard reaction conditions µ The amount of enzyme required for mol phosphate was defined as 1 active unit.Preservation system20 mM Tris-HCl; 100 mM NaCl; 1 mM DTT; 0.1 mM EDTA; 50% (v/v) Glycerol; pH 8.0。 Storage temperature-20±5 ℃。Matters needing attention1. the enzyme has activity in various reaction buffers. Generally, the enzyme can be directly added in HDA, lamp and other experiments.2. the dosage of the enzyme needs to be optimized in different experiments, usually adjusted at the concentration of 0.05~1u/ml.3. the optimum reaction temperature of the enzyme was 25 ℃, and it was active at 16~37 ℃, and the enzyme could be inactivated at 65 ℃ for 10min.4. cofactor: mg2+ is necessary for enzyme activity... Read More | Purity>95% (SDS-PAGE) Endotoxin level<1.0 EU/µgFunctionInhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells | Purity> 97% (SDS-PAGE&HPLC)Endotoxin level<0.1 EU/µgFunctionProduced by macrophages, IFN-alpha have antiviral activities. Interferon stimulates the production of two enzymes: a protein kinase and an oligoadenylate synthetase |