| Description | Products contentS665634Component1 mL5 mLStorageS665634A2×Super Pfx Master Mix1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle.S665634BddH₂O1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle. Products IntroductionIt is an optimized 2× premixed reagent consisting of Super Pfx DNA Products contentS665634Component1 mL5 mLStorageS665634A2×Super Pfx Master Mix1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle.S665634BddH₂O1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle. Products IntroductionIt is an optimized 2× premixed reagent consisting of Super Pfx DNA Polymerase, Mg2+, dNTPs, and reaction buffer, and requires only the addition of primers and DNA templates for the PCR reaction.Super Pfx DNA Polymerase has a 3´→5´ exonuclease activity, and is approximately 100 times more fidelity than Taq DNA Polymerase. Super Pfx DNA Polymerase has 3´→5´ exonuclease activity and is about 100 times more fidelity than Taq DNA Polymerase, making it ideal for cloning. With the introduction of elongation enhancement technology, this enzyme enables high-speed PCR with an elongation speed of 10 sec/kb and an amplification length of up to 20 kb, while maintaining high fidelity. In addition, this product combines a high success rate with the ability to amplify long fragments that are difficult to amplify, GC-rich regions, and templates that are too large to be amplified. The amplified PCR product does not have an "A" base at the 3´ end and can be directly cloned into a flat-end vector. This product is divided into the standard 2×Master Mix without Loading Dye and the 2×Master Mix with Loading Dye.Product Characteristics1. High-speed PCR possibleThe extension time can be set to 10sec for amplification of target fragments of less than 1kb, and 20-30sec/kb for amplification of target fragments of 1-10kb, which significantly reduces the PCR reaction time compared to conventional reagents.2. High fidelityThe fidelity of Taq DNA polymerase is about 100 times higher than that of Taq DNA polymerase, and long target fragments can be amplified quickly and with high fidelity, and the amplified products can be used for a variety of purposes.3. simple and convenientThis reagent contains all PCR components except primers and templates, which facilitates the operation and improves the reproducibility of the results. In addition, 2×Super Pfx Master Mix (Dye) contains Loading Dye, which can be used for gel electrophoresis directly after the reaction.caveatPrimers and templates with uracil are not suitable for this product.Instructions for usePCR Reactors1. Before preparing the reaction solution, please melt and mix the reagents completely before use.After adding the components, please mix them well and transfer them to the PCR instrument quickly.・The recommended final primer concentration is 0.4-0.5 µM, but for amplification of long fragments of 10 kb or more, a final primer concentration of 0.3 µM can increase the amount of amplified product.PCR reaction program1. Before preparing the reaction solution, please melt and mix the reagents completely before use.take note of 1）Pre-denaturation: for most purified templates, 98℃, 30sec is sufficient; for complex templates, the pre-denaturation time can be extended, not more than 3min. 2）Annealing: In general, the annealing temperature is 3-5°C lower than the primer Tm. When a non-specific reaction occurs, increase the annealing temperature appropriately. If necessary, a temperature gradient can be established to find the optimal temperature for primer-template binding. For high Tm primers, a two-step cycle can be used, combining annealing and extension into one step.3）Extension: for complex genomic samples, extension times are typically 20-30sec/kb, but can be reduced to 10sec/kb for simple templates (plasmids, E. coli, etc.) or complex templates <1kb. cDNAs, or complex templates >6kb, can be increased to 40-50sec/kb, if desired... Read More | Protein Purity>95% by SDS-PAGEExtinction Coeff.A276 nm = 0.456 at 1.0 mg/mLMolecular Weight8,759 Da (single chain)General DescriptionNatural human C4a is prepared by cleavage of human C4 protein by human C1s. It is produced during activation of both the classical and lectin pathways of complementProtein Purity>95% by SDS-PAGEExtinction Coeff.A276 nm = 0.456 at 1.0 mg/mLMolecular Weight8,759 Da (single chain)General DescriptionNatural human C4a is prepared by cleavage of human C4 protein by human C1s. It is produced during activation of both the classical and lectin pathways of complement. C4a is a member of the anaphylatoxin family of three proteins (C3a, C4a and C5a) produced by the activation of complement (Hugli, T.E. et al. (1981)). It is an unglycosylated polypeptidecontaining 77 amino acids with a molecular mass of 8,759 daltons. Many of the biological functions of C4a are similar to those of C3a, but the specific activities are far below those of C3a. C4a activity is so low, in fact, that it was initially thought to be inactive. These measured activities include inducing muscle contraction in the guinea pig ileum test (spasmogenic activity), desensitization of muscle to C3a stimulation suggesting that the same receptor for both C3a and C4a is involved (tachyphylactic activity) and inducing vascular permeability in human skin (Gorski J.P. et al. (1979)). C4a does not show tachyphylactic activity against C5a or chemotactic activity. Removal of the C-terminal arginine by serum carboxypeptidase N destroys all these activities (Meuller-Ortiz, S.L., et al. (2009)). C4a appears to act through the C3a receptor (C3aR) which is a G-protein coupled receptor found widely distributed on peripheral tissues, lymphoid cells (neutrohphils, monocyes, and eosinophils) and in the central nervous system (astrocytes, neurons and glial cells) (Law, S.K.A. and Reid, K.B.M. (1995)). Physical Characteristics & StructureMolecular weight: 8,759 calculated molecular mass. Observed mass (MALDI-TOF) is 8,762 + 9 mass units. pI = 9.0 to 9.5 (Gorski, J.P. et al. (1981))Amino acid sequence (77 amino acids): NVNFQKAINE KLGQYASPTA KRCCQDGVTR LPMMRSCEQR AARVQQPDCR EPFLSCCQFA ESLRKKSRDK GQAGLQRC4a is thought to be structurally very similar to C3a and C5a to which it is homologous. Thus its 3D structure is probably similar to the X-ray-derived crystal structureof C3a (Huber, R. et al. (1980)) and the NMR derived structure of C3a: Nettesheim, D.G. et al. (1988); Murray, I. et al. (1999).FunctionSee General Description above. C4a exhibits much weaker biological activities than C3a and C5a. Its activity in inducing erythema and edema in human skin is 25,000-fold weaker than that of C5a and 100-fold weaker than C3a per nanomole. The spasmogenic activity of C4a is 2000-fold weaker than C5a and 100-fold weaker than that of C3a. Due to these differences the role of C4a in these responses in vivo is thought to be negligible.AssaysTwo well established assays for C4a and C3a functional activities include induction of contraction in the guinea pig ileum and the permeation of a dye such as trypan blue from the vasculature into skin. The anaphylatoxins also induce 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. The other assays used for C3a (Dodds, A.W. and Sim, R.B. (1997)) should also respond to C4a, but few reports have described utilizing these assays with C4a. ELISA kits for the assay of C4a levels (or more correctly C4a desArg levels) in blood and other fluids are sold by several companies. 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 significant levels of all three anaphylatoxins. Although these may represent the resting concentration in vivo it is difficult to draw or store blood without some complement 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 C4 in blood (600µg/mL) would result in ~3,400 nM C4a (~30 µg/mL). Due to the low biological activity of C4a it could require activation of most of the C4 in a small region to achieve the micromolar C4a concentrations necessary to elicit a response.RegulationC4a levels are regulated by three processes: formation, inactivation and clearance. There are two enzymes that cleave C4 and release C4a: C1s and MASP-2. C4a is “inactivated” by removal of its C-terminal arginine amino acid. The product C4a desArg (or C4a 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 C4a is converted to C4a desArg within minutes of its formation. Inactivated C4a lack measurable biological activity. Because of the large number of cells bearing C3a/C4areceptors (endothelial, immune, smooth muscle, neuronal, etc.) the capture, internalization and digestion of C4a and C4a desArg probably results in its removal from circulation.DeficienciesA deficiency of C4 or a deficiency of all of the enzymes that cleave C4 to generate C4a could result in the absence of C4a. There are no known complete deficiencies of all ofthe C4 cleaving enzymes. Examples of C4 deficient humans and mice exist (Wessels, M.R. et al. (1995)), but the degree to which pathologies associated with C4 deficiency are due to the lack of C4 or the absence of C4a is unclear. DiseasesThere are no known diseases connected to C4a or C4a desArg. Precautions/Toxicity/HazardsThe source of C4a is human serum, therefore appropriate precautions must be observed 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.Injection can cause anaphylatic shock which is a generalized circulatory collapse similar to that caused by an allergic reaction.Hazard Code: B WGK Germany 3... Read More | Product introduction：The additive contains a variety of cytokines, human albumin and other components. In order to ensure the activity, it is recommended that the additive part be thawed once and then sub packaged and frozen again. It is not advisable to freeze and thaw repeatedly. The Product introduction：The additive contains a variety of cytokines, human albumin and other components. In order to ensure the activity, it is recommended that the additive part be thawed once and then sub packaged and frozen again. It is not advisable to freeze and thaw repeatedly. The product contains no animal serum or animal serum components, no animal protein components, and no antibiotics.Matters needing attention：1. try to reduce the number of repeated freezing and thawing to avoid efficiency decline. 2. it is not suitable to place it at room temperature for a long time. 3. pay attention to aseptic operation and try to avoid pollution. 4. please wear experimental clothes and disposable gloves for operationScope of application：Cell culture additives... Read More | Purity>95% (SDS-PAGE&HPLC) Endotoxin level<0.1 EU/µgFunctionMay regulate apoptosis, cell proliferation and cell differentiation. Binds beta-galactoside and a wide array of complex carbohydrates. Inhibits CD45 protein phosphatase activity and therefore the dephosphorylation of Lyn Purity>95% (SDS-PAGE&HPLC) Endotoxin level<0.1 EU/µgFunctionMay regulate apoptosis, cell proliferation and cell differentiation. Binds beta-galactoside and a wide array of complex carbohydrates. Inhibits CD45 protein phosphatase activity and therefore the dephosphorylation of Lyn kinase.Gal-1 is also engaged in many protein-protein interactions. Gal-1 plays a number of crucial roles in neuronal cell differentiation and survival in both the central and the peripheral nervous systems, and the establishment and maintenance of T-cell tolerance and homeostasis in vivo... Read More | Inquire |