| Description | Extinction CoeffA280 nm = 1.15 at 1.0 mg/mL for pure C1sMolecular Weight92,000 Da (1 chain)Protein Purity>90 % by SDS PAGEGeneral Description C1r proenzyme is a single chain 92,000 dalton protein that is the native form of C1r enzyme. C1r is a subunit of the C1 complex which is the first Extinction CoeffA280 nm = 1.15 at 1.0 mg/mL for pure C1sMolecular Weight92,000 Da (1 chain)Protein Purity>90 % by SDS PAGEGeneral Description C1r proenzyme is a single chain 92,000 dalton protein that is the native form of C1r enzyme. C1r is a subunit of the C1 complex which is the first complement component in the cascade referred to as the classical pathway of complement. C1r proenzyme is an inactive zymogen until C1 is activated. C1r is activated when C1 binds to and is activated by antibodies bound to antigens (immune complexes) yielding C1r enzyme, the first protease that initiates the cascade. C1 is a non-covalent calciumdependent complex of one C1q, two C1r and two C1s molecules. Each C1q binds through two or more of its six arms to the Fc domains of IgG or IgM. The binding of multiple arms to immune complexes causes the two C1r proteins in the complex (protease zymogens) to activate producing two proteases that cleave and activate the two C1s protease zymogens in the complex (Morikis, D. and Lambris, J.D. (2005)). The activation of C1r results from cleavage of C1r into two fragments of 57,000 and 35,000 daltons. Activation of the bound C1s molecule is the only known function of C1r enzyme. Activated C1s cleaves complement component C4 releasing C4a and initiating covalent attachment of C4b to the activating surface. Activated C1s also cleaves C2 and the larger fragment of C2 binds to the surface-attached C4b forming C4b,C2a the C3/C5 convertase of the classical pathway. Physical Characteristics & Structure C1r proenzyme is a 92,000 dalton, single chain peptide that exhibits trypsin-like proteolytic specificity for arginyl bonds present in its two natural substrates, C1s and itself. C1r is present in plasma at 34 ug/mL (0.2µM) (Cooper, 1985). C1r proenzyme is an unstable zymogen and it spontaneously activates (Dodds, A.W. and Sim, R.B. editors (1997); Morikis, D. and Lambris, J.D. editors. (2005)) by cleaving a peptide bond in C1r producing a 57,000 dalton heavy chain and a 35,000 dalton light chain. This is the form sold as C1r enzyme. This self-activation occurs rapidly in the C1 complex upon binding to an immune complex and it occurs slowly with pure C1r. Two C1r form a C1r-C1r complex in the presence of calcium which in turn forms a stable complex with two C1s molecules in the presence of calcium. This tetramer can exist in solution, but in the presence of C1q it binds to C1q forming the C1 complex, which is stable in the presence of calcium. C1r self-activation is controlled in part by a weak association with C1esterase inhibitor (C1-INH) when it is in the C1 complex and similar stabilization occurs with purified C1r (Ziccardi, R.J. (1982)). C1r enzyme, however, is irreversibly inactivated by binding to C1-INH. Function The biological functions of C1r proenzyme are described above in the General Description and Physical Characteristics sections. C1r proenzyme can be used in the presence of calcium to form the C1 complex with C1q and C1s proenzyme. The C1r proenzyme will rapidly self-activate to C1r enzyme when the C1q in the C1 complex binds to immune complexes such as EA cells bearing antibodies. EA are sheep erythrocytes with rabbit IgM anti-sheep erythrocytes antibodies bound to their surface(Morgan, B.P. ed. (2000)). The activated C1r enzyme will rapidly activate the two C1s proenzymes to form C1s enzymes and the resulting C1q-C1r2-C1s2 complex is a fully active C1 molecule which will activate C4 and C2 (Dodds, A.W. and Sim, R.B. editors (1997); Morgan, B.P. ed. (2000)). AssaysThe unit of classical pathway activity is the CH50. A similar unit, the C1rH50, can be used to quantitate the activity of C1r proenzyme for which we have developed a hemolytic assay. The C1rH50 assay uses our newly developed C1r-Dpl and measures the lysis of EA (classical pathway) as a function of the concentration of added test sample or standard purified C1r proenzyme. A C1rH50 unit is the amount of functional C1r proenzyme needed to lyse 50% of 3 x 10⁷EA cells (antibodysensitized sheep erythrocytes when that amount of C1r proenzyme is incubated with the recommended volume of C1r-Dpl in GVB⁺⁺ in a total volume of 500 µL for 30 min at 37℃. This amount of C1r proenzyme indicates the sensitivity of the assay for C1r proenzyme which is typically less than 10 ng C1r proenzyme with 10 µL C1r-Dpl. See the Certificate of Analysis for lot specific values.ApplicationsSee sections titled Function and Assays above. Regulation Activated C1r is rapidly inactivated by C1-INH. The spontaneous activation of C1r observed with pure C1 and pure C1r proenzyme is minimized by the presence of C1- INH which rapidly inactivates spontaneously activated C1r enzyme. Stabilization of the proenzyme is also due to existence of a weak complex between C1-INH and C1r proenzyme. This association apparently stabilizes C1 thus preventing spontaneous activation in serum (Ziccardi, R.J. (1982)). Separation of C1-INH from C1 during purification is one of the reasons that isolated C1 and C1r proenzyme are unstable and prone to spontaneous activation. Genetics The EMBL/Genbank cDNA accession number for C1r is M14058. The genes for C1r and C1s are closely linked and located on chromosome 12p13. DeficienciesDeficiencies of each of the three components of C1 have been found (Ross, G.D. (1986)). C1r and C1s deficient patients are prone to systemic lupus erythematosus (SLE) and recurrent pyogenic infections (Rother, K., et al. (1998)). They lack classical pathway function and may or may not exhibit C1r antigen in blood. DiseasesSee section titled Deficiencies above. 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... Read More | Product Content:F665667Component5 mL40 mLStorageF665667A2×Flash PCR MasterMix (Dye) 5×1 mL 40×1 mL-20℃. Avoid freeze/thaw cycle.F665667BddH2O 5×1 mL40×1 mL-20℃. Avoid freeze/thaw cycle. Products Introduction This product is a premixed system consisting of a new Product Content:F665667Component5 mL40 mLStorageF665667A2×Flash PCR MasterMix (Dye) 5×1 mL 40×1 mL-20℃. Avoid freeze/thaw cycle.F665667BddH2O 5×1 mL40×1 mL-20℃. Avoid freeze/thaw cycle. Products Introduction This product is a premixed system consisting of a new high efficient fast DNA Polymerase, Mg2+, dNTPs, and PCR stabilizers and enhancers at 2× concentration. It is a new rapid DNA polymerase developed by CombiSigma with high amplification speed and stability. The extension speed is up to 5 s/kb, and the PCR can be completed in as little as 15 minutes, while longer fragments (>3 kb) or complex templates can be extended at a speed of 10-30 s/kb or a higher number of cycles. The unique MasterMix formula makes the whole reaction system very stable, while complex templates can be amplified effectively, and more than 98% of PCR amplification can be successful in one run. Simply add the DNA template and primers and top up with water to minimize human error, contamination and time.The dye (blue) has been added to the product and it is ready for electrophoretic detection at the end of the reaction. The PCR product is amplified with an 'A' base at the 3′ end and can therefore be used directly for T/A cloning and is suitable for use in the CombiVerge Seamless Cloning Kit, T4 Ligation Kit and sensory products.This product is mainly suitable for ultra-fast PCR, complex templates, complex secondary structures, gene cloning and large-scale genetic testing that requires high fidelity. quality control No exogenous nuclease activity was detected; no host residual DNA was detected by PCR; single-copy genes in various genomes could be amplified efficiently. UsageThe following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using 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.PCR reaction system Note: 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.PCR reaction conditions Note: 1) Note: For simple templates, the pre-denaturation time can be controlled at 30 s-1 min, for complex templates such as bacterial fluids, the pre-denaturation time can be increased to 2 min.Optimization of parameter settings 1. Template DNA amount setting:Excessive amounts of template may result in non-specific amplification or smear. The recommended amount of template DNA in a 50 µl PCR reaction system is as follows:-Human genomic DNA 5 ng-500 ng-Escherichia coli genomic DNA 50 pg-100 ng-plasmid DNA 10 pg-1 ng 1. 30-35 number of cycles2. Primer concentration setting: The primer concentration can be set between 0.1 µM and 1.0 µM. A low primer concentration may result in low amplification products. Too high a primer concentration will inhibit specific amplification and may result in non-specific amplification.3. Annealing temperature setting: In general, the annealing temperature is 5℃ lower than the melting temperature of amplification primer Tm, so the annealing temperature can be lowered appropriately when the desired amplification efficiency cannot be obtained; the annealing temperature can be raised appropriately when non-specific reaction occurs. For complex templates, it is necessary to adjust the annealing temperature to achieve efficient amplification.4. Extension time setting: The extension time should be set according to the size of the amplified fragments. The following extension times are recommended: simple templates such as plasmids: 5-15 s/kb; regular genomes, cDNA templates: 10-15 s/kb; complex templates, crude templates: 20-30 s/kb; (the extension time should not be too short and should be at least 5 s/kb, but should not exceed 30 s/kb).5. Number of cycles: 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 | Gly-Pro-pNA hydrochloride is a dipeptidyl peptidase inhibitor that inhibits dipeptidyl peptidase II, dipeptidyl peptidase IV and dipeptidyl peptidase IX | Inquire | Recombinant Human Serum Albumin (rHSA) is an active compound and possesses an identical conformation to plasma derived HSA. Recombinant Human Serum Albumin (rHSA) has no difference between rHSA and plasma derived HSA. Recombinant Human Serum Albumin (rHSAAppearance:SolidBiological Activity:Recombinant Human Serum Albumin (rHSA) is an active compound and possesses an identical conformation to plasma derived HSA. Recombinant Human Serum Albumin (rHSA) has no difference between rHSA and plasma derived HSA. Recombinant Human Serum Albumin (rHSAAppearance:SolidBiological Activity:Recombinant Human Serum Albumin (rHSA) is an active compound and possesses an identical conformation to plasma derived HSA. Recombinant Human Serum Albumin (rHSA) has no difference between rHSA and plasma derived HSA. Recombinant Human Serum Albumin (rHSA... Read More |