| Description | Products contentS665683Component1mL5mLStorageS665683A2×Super Kfx MasterMix1 mL5×1 mL -20℃. Avoid freeze/thaw cycle.S665683BddH2O1 mL5×1 mL -20℃. Avoid freeze/thaw cycle.Products IntroductionThis product is a premixed system consisting of Super Kfx DNA Polymerase, Mg2+, Products contentS665683Component1mL5mLStorageS665683A2×Super Kfx MasterMix1 mL5×1 mL -20℃. Avoid freeze/thaw cycle.S665683BddH2O1 mL5×1 mL -20℃. Avoid freeze/thaw cycle.Products IntroductionThis product is a premixed system consisting of Super Kfx DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers at a concentration of 2×. Super Kfx DNA Polymerase is a fast, high-fidelity DNA polymerase with high amplification efficiency, which possesses 5′-3′DNA polymerase activity and 3′-5′ exonuclease activity. With 5′-3′ DNA polymerase activity and 3′-5′ exonuclease activity, the enzyme has the advantages of strong amplification ability, high fidelity and high specificity, etc. 2×Mix has added unique amplification enhancement factors and extension factors, and the unique formula makes the whole reaction system very stable and easy to operate, which is suitable for the amplification of various fragments and templates. The product is suitable for gene cloning, second generation library amplification, targeted gene mutation, SNP amplification experiments. quality controlNo exogenous nuclease activity, can effectively amplify various templates; stored at 2-8℃ for one month, no obvious activity change.UsageThe following examples are conventional PCR reaction systems and conditions, which should be improved and optimized according to the template, primer structure and target fragment size.1. PCR reaction system All operations should be carried out on ice, please mix the components thoroughly after thawing, after use, please promptly return to -20 ℃ for storage. 2. PCR reaction system take note of1）Priority is given to three-step amplification; if the three-step method fails to amplify the target product or if the primer Tm value is greater than 68°C, try the two-step method.2）Denaturation: pre-denaturation of simple templates 98°C, 30s-1min, for complex templates, the pre-denaturation time can be extended to 3min. 3）Annealing: In general, the annealing temperature is 3-5℃ lower than the Tm value of the primers. If the desired amplification efficiency cannot be obtained, the annealing temperature should be changed in a gradient to optimize the results; if a non-specific reaction occurs, the annealing temperature should be increased appropriately.4）Extension: The extension time should be set according to the length of the amplified fragments and the complexity of the template, the amplification efficiency of this product is 4-6kb/min, for long fragments and templates with high complexity it is recommended that 2-4kb/min. 5）Cycling times: the number of cycles can be set according to the downstream application of the amplified product. If the number of cycles is too small, the amplification will be insufficient, and if the number of cycles is too large, the chance of mismatch will be increased, so the number of cycles should be minimized under the premise of guaranteeing the yield of the product... Read More | Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 10.16 at 1.0 mg/ml for pure C3Molecular Weight187,000 Da (2 chains)General DescriptionRat C3 is purified from pooled normal rat serum. C3 is central to the activation of all three pathways of complement activation (Law, S.K.A. and Reid, KProtein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 10.16 at 1.0 mg/ml for pure C3Molecular Weight187,000 Da (2 chains)General DescriptionRat C3 is purified from pooled normal rat 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 & StructureThe calculated molecular weight of rat C3 based on its amino acid sequence is 184,111daltons (without the signal peptide) and is similar to that of human C3 (185,000 daltons).The molecular weight of rat C3 as determined by SDS/polyacrylamide gel electrophoresis has been reported by Daha, M.R. et al., (1979) to be 187,000 daltons composed of two disulfide linked chains, alpha chain (123,000 daltons) and beta chain (76,000 daltons). The extinction coefficient of rat C3 (E1%/280nm = 10.16) is calculated based on its amino acid sequence using ProtParam and assumes all pairs of Cys residues form cystines (i.e. a pair of cysteine molecules are joined by a disulfide bond). The theoretical pI of rat C3 is 6.12. The normal plasma concentration of C3 inWistar rats has been reported to be 0.581mg/ml (Daha, M.R. et al., (1979)).FunctionThe biological functions of C3 are described above in the General Description section.GeneticsRat C3 chromosome location 9. The NCBI Gene ID number for rat C3 is 24232 and UniProt accession number is P01026.Precautions/Toxicity/HazardsThis protein is purified from animal plasma/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.Daha MR, Stuffers-Heiman M, Kijlstra A and Van ES LA. (1979) Isolation and characterization of the third component of rat complement. Immunology 36:63-70... Read More | 1、Product attributeReaction time：short (up to 20 minutes) at 20-37°CLot-to-lot variation：<5%Boiling point : 100℃pH-Value (at 20 °C): 3.5-4.0Density (20℃) : 1.0111 g/cm³Appearance: colourless to pale blue liquidOdour: odourlessRecommend Incubation 1、Product attributeReaction time：short (up to 20 minutes) at 20-37°CLot-to-lot variation：<5%Boiling point : 100℃pH-Value (at 20 °C): 3.5-4.0Density (20℃) : 1.0111 g/cm³Appearance: colourless to pale blue liquidOdour: odourlessRecommend Incubation temperature: 20-37 °C2、Requirements for storage rooms and vessels1.Keep container tightly closed.2.Keep cool. protected from light3.Keep/Store only in original container.4.Never return spills in original containers for reuse.5. Keep away from: Food and feeding stuffs3、It is a ready-to-use, labelling-free TMB-substrate solution.4、Biosafety informationThis mixture is not classified as hazardous in accordance with Regulation (EC) No 1272/2008；5、Advantage1. Very high absorbance yield2. Very low background signals3. Certified long-term stability4. Regeneration following light exposure... Read More | Inquire | Inquire |