| Description | Products contentNote: The 10 x PCR Buffer for this product contains 15 mM magnesium ions. Products IntroductionFastPrime DNA Polymerase is a mixture of anti-Taq enzyme monoclonal antibody and Taq DNA Polymerase with high amplification and fidelity properties for HOT Start PCR. When using FastPrime Products contentNote: The 10 x PCR Buffer for this product contains 15 mM magnesium ions. Products IntroductionFastPrime DNA Polymerase is a mixture of anti-Taq enzyme monoclonal antibody and Taq DNA Polymerase with high amplification and fidelity properties for HOT Start PCR. When using FastPrime DNA Polymerase for PCR amplification, Taq enzyme antibody binds to Taq enzyme and inhibits DNA polymerase activity before denaturation at low temperature. When using FastPrime DNA Polymerase for PCR amplification, before denaturation at high temperature, Taqase antibody binds to Taqase to inhibit DNA polymerase activity, which can effectively inhibit the non-specific annealing of primers and non-specific amplification caused by primer dimer at low temperature, and the Taqase antibody denatures during the initial DNA denaturation step of the PCR reaction, and the activity of DNA polymerase is restored, thus achieving the effect of Hot Start. No special inactivation of the Taqase antibody is required for the use of this product, and it can be used under conventional PCR reaction conditions.FastPrime DNA Polymerase has 5′→3′ DNA polymerase activity, 5′→3′ exonuclease and 3′→5′ exonuclease activity. Compared with Taq DNA Polymerase, FastPrime DNA Polymerase has high amplification efficiency and low mismatch rate, and can amplify DNA fragments efficiently. The PCR products amplified with this product have an "A" base at the 3′ end, which can be directly used for T/A cloning. This product is suitable for routine PCR reactions and gene cloning reactions that require high fidelity.Definition of activityThe amount of enzyme required to dope 10 nmol of deoxyribonucleotide into acid-insoluble material was defined as 1 activity unit (U) at 74°C for 30 min, using activated salmon sperm DNA as template/primer.quality control After several column purification, its purity is more than 99% by SDS-PAGE; no exogenous nuclease activity is detected; no host residual DNA is detected by PCR method; it can effectively amplify single-copy genes in the human genome.UsageThe following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment of human genomic DNA as a template, which should be improved and optimized according to the template, primer structure and size of the target fragment in actual operation.1. PCR reaction systemNote: Please use the final concentration of 0.1-1.0 µM as a reference for setting the range of primer concentration. If the amplification efficiency is not high, the primer concentration can be increased; if a non-specific reaction occurs, the primer concentration can be decreased to optimize the reaction system. 2.PCR reaction conditionsAttention: 1）In general, the annealing temperature is 5°C lower than the melting temperature of the amplification primer, Tm. When the desired amplification efficiency cannot be obtained, the annealing temperature should be lowered appropriately; when non-specific reaction occurs, the annealing temperature should be increased, so as to optimize the reaction conditions.2）The extension time should be set according to the size of the amplified fragment, and the amplification efficiency of this product is 2 kb/min. 3) The number of cycles can be set according to the downstream application of the amplified product. If the number of cycles is too low, the amount of amplification will be insufficient; if the number of cycles is too high, the chance of mismatch will increase and the non-specific background will be serious. Therefore, the number of cycles should be minimized under the premise of ensuring the product yield... Read More | Inquire | Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 0.974 at 1.0 mg/ml for pure C3bMolecular Weight185,000 Da (2 chains)General DescriptionCynomolgus monkey C3 (cyno C3) is purified from pooled normal cynomolgus monkey serum. C3 is central to the activation of all three pathways of Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 0.974 at 1.0 mg/ml for pure C3bMolecular Weight185,000 Da (2 chains)General DescriptionCynomolgus monkey C3 (cyno C3) is purified from pooled normal cynomolgus monkey serum. C3 is central to the activation of all three pathways of complement activation (Law, S.K.A. and Reid, K.B.M. (1995)). Initiation of each pathway generates proteolytic enzyme complexes (C3 convertases) which are bound to the target surface. These enzymes cleave a peptide bond in C3 releasing the anaphylatoxin C3a and activating C3b. For a brief time (~60 µs) this nascent C3b is capable of reacting with and covalently coupling to hydroxyl groups on the target surface. Carbohydrates are the favored target, but protein hydroxyls and amino groups also react. This process of tagging the target surface with C3b is called opsonization. The reactive site in nascent C3b is a thioester (Tack B.J., et al. (1980); Pangburn M.K. and MüllerEberhard H.J. (1980)) and C3b is linked to the target through a covalent ester bond (an amide bond is formed if C3b is attached to amino groups). Most of the C3 activated during complement activation never attaches to the surface because its thioester reacts with water forming fluid phase C3b which is rapidly inactivated by factors H and I forming iC3b. Surface-bound C3b is necessary in all three pathways for efficient activation of C5 and formation of C5b-9 complexes that lyse the target cell membrane. Surface-bound C3b and its breakdown products iC3b and C3d are recognized by numerous receptors on lymphoid and phagocytic cells which use the C3b ligand to stimulate antigen presentation to cells of the adaptive immune system. The end result is an expansion of target-specific B-cell and T-cell populations.Physical Characteristics & StructureCynomolgus monkey C3 is an uncharacterized protein. The calculated molecular weight based on its amino acid sequence is 184,926 daltons similar to that of human C3 (185,000 daltons). Like human C3, cyno C3 is composed of two disulfide-linked chains. Analysis of purified cyno C3 by SDS/polyacrylamide gel electrophoresis under non-reduced conditions shows the mobility of cyno C3 to be similar to that of human C3. Under reduced conditions, the migration of the alpha chain of cyno C3 is comparable to that of human C3 alpha chain (110,000 daltons) while the beta chain migrates slightly ahead of the human C3 beta chain (75,000daltons).The extinction coefficient of cyno C3 is calculated from its amino acid sequence using ProtParam and assumes all pairs of Cys residues form cystines (i.e. a pair of cystine molecules are joined by a disulfide bond). The theoretical pI value for cyno monkey C3 is 6.03. Employing immunoturbidimetric method the serum concentration of cyno C3 has been reported to be 1.27 mg/ml in males and 1.1 mg/ml in female monkeys (Park H-K et al., (2016)). FunctionThe biological functions of C3 are described above in the General Description and Physical Characteristics sections.GeneticsCynomolgus monkey C3 chromosome location 19. The NCBI Gene ID number for Cynomolgus monkey C3 is 102131458 and UniProt accession number is A0A2K5VPN1.Precautions/Toxicity/HazardsThis protein is purified from animal serum and therefore precautions appropriate for handling any animal blood-derived product must be used.ReferencesLaw, S.K.A. and Reid, K.B.M. (1995) Complement 2nd Edition (ISBN 0199633568) Oxford University Press, Oxford.Tack BF, Harrison RA, Janatova J, Thomas ML, Prahl JW. (1980) Evidence for presence of an internal thiolester bond in third component of human complement. Proc Natl Acad Sci U S A. 77:5764-8.Pangburn M.K. and Müller-Eberhard H.J. (1980) Relation of putative thioester bond in C3 to activation of the alternative pathway and the binding of C3b to biological targets of complement. J Exp Med. 152:1102-14.Park H-K, Cho J-W, Lee B-S, Park H, Han J-S, Yang M-J, Im W-J, Park D-Y, Kim W-J, Han SC, Kim Y-B. (2016) Reference values of clinical pathology parameters in cynomolgus monkeys (Macaca fascicularis) used in preclinical studies. Lab Anim Res. 32(2):79-86... Read More | Inquire | Protein:BovineEnzyme:Horseradish peroxidase |