| Description | Aladdin's 6% SDS-PAGE Resolving Gel Master Mix contains almost all reagents required for the preparation of 6% SDS-PAGE resolving gels, except for polymerization catalysts such as ammonium persulfate and TEMED.Aladdin's SDS-PAGE Resolving Gel Master Mix series can be used to prepare 5 common Aladdin's 6% SDS-PAGE Resolving Gel Master Mix contains almost all reagents required for the preparation of 6% SDS-PAGE resolving gels, except for polymerization catalysts such as ammonium persulfate and TEMED.Aladdin's SDS-PAGE Resolving Gel Master Mix series can be used to prepare 5 common concentrations of gelsPrecautions：Ammonium Persulfate or its substitute (, ST005) and TEMED are required but not provided in this product.This product contains Acr-Bis which is potentially neurotoxic. Please take effective measures to avoid direct contact with the human body or inhalation.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1.According to the size of a target protein, decide an appropriate concentration of SDS-PAGE resolving gel. Please refer to the table below.SDS-PAGE Resolving Gel Concentration Optimal Separation Range6%50-150kD8%30-90kD10 -80kD12%12-60kD15%10-40kD2.Prepare a 10% ammonium persulfate solution with ddH2O or other high-purity water. Solution of ammonium persulfate or its substitute is prone to failure and should be prepared freshly or kept frozen for multiple uses.3.Prepare the resolving gel according to the table below. For example, add 100µl of 10% ammonium persulfate and 4µl of TEMED into 10ml of 6% SDS-PAGE Resolving Gel Master Mix, mix well, and immediately pour into gel cassette. Overlay with the top layer of water-saturated butanol, isopropanol, 0.1% SDS or distilled water. Leave at room temperature (~25℃) until fully solidified (usually within 10-30 minutes). Note: The amount of polymerization catalyst in the table below is recommended for polymerization at 25℃. It can be adjusted appropriately according to the temperature. For example, when the room temperature is lower than 20℃, add more TEMED and 10% ammonium persulfate appropriately to promote gel solidification. ReagentsVolume of each component (ml) required for different volumes of SDS-PAGE resolving gel6% SDS-PAGE Resolving Gel Master Mix5101520305010% ammonium persulfate or its substitute0.050.10.150.20.30.5TEMED 0.0020.0040.0060.0080.0120.024.After complete polymerization of the resolving gel, remove the liquid used for sealing, and then prepare the SDS-PAGE stacking gel with 's SDS-PAGE Stacking Gel Master Mix. 5.If the prepared gel is not to be used on the same day, it can be stored at 4℃ for 1-2 days... Read More | Inquire | Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 0.631 at 1.0 mg/ml for pure C1qMolecular Weight400,000 Da (18 chains)General DescriptionRat C1q is purified from pooled normal rat serum. C1q is part of the C1 complex, which is the first complement component in the classical pathway of Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 0.631 at 1.0 mg/ml for pure C1qMolecular Weight400,000 Da (18 chains)General DescriptionRat C1q is purified from pooled normal rat serum. C1q is part of the C1 complex, which is the first complement component in the classical pathway of complement. The C1 complex is a non-covalent assembly of three different proteins (C1q, C1r, and C1s) bound together in a calcium-dependent complex. C1q has six extended arms with domains at the end of each arm that bind to the Fc domains of immunoglobulins such as IgG or IgM. When antibodies bind toantigens, forming immune complexes, they cluster allowing two or more of the six C1q arms to bind to the Fc domains of antibodies. Rat IgG2 is very efficient when compared to IgG1 in activating complement (Medgyesi, G.A et., al., 1981). This is in contrast to the human system in which IgG1 activates complement but not IgG2 (Redpath, S. et. al., 1998). The binding of multiple arms of C1q to immune complexes causes the two C1r proteins in the complex (protease zymogens) to auto-activate. The activated C1r proteases cleave and activate the two C1s protease zymogens in the complex. The 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.Rat IgG1 cannot activate complement whereas rat IgG2 does.Physical Characteristics & StructureThe apparent molecular weight of rat C1q as determined by gel filtration has been reported to be 400,000 by Veerhuis, R. et al., (1985) and is calculated to be 420,000 based on its amino acid sequence. Rat C1q is a high molecular weight complex of 18 polypeptide chains. Each of the six arms of rat C1q contains three chains, an A chain (~30,000 daltons), a B chain (~28,000 daltons) and a C chain (~26,000 daltons) as determined by SDS/polyacrylamide gel electrophoresis (Wing, M.G. et al., (1993)).FunctionThe biological functions of C1q are described above in the General Description and Physical Characteristics sections.ApplicationsRat C1q can be used to coat ELISA plates to capture and quantitate immune complexes in samples from rat models used for studying immune complex related diseases and conditions.GeneticsNCBI Gene ID numbers for rat C1q are: C1q A chain (298566), C1q B chain (29687), and C1q C chain (362634). The genes for C1q chains A, B and C are all located on chromosome 5. The UniprotKB primary accession numbers for rat C1q are: C1q A chain (P31720), C1q B chain (P31721), and C1q C chain (P31722).Precautions/Toxicity/HazardsThis protein is purified from animal plasma/serum and therefore precautions appropriate for handling any animal blood-derived product must be used.ReferencesMedgyesi, G.A et., Miklos, K., Kulics, J., Fust, G., and Gergely, J. Bazin, H. (1981). Classes and subclasses of rat antibodies: reaction with the antigen and interaction of the complex with the complement system. Immunology 43, 171-176.Redpath, S., Michaelsen, T., Sandlie, I. and Clark, M. R. (1998). Activation of complement by human IgG1 and human IgG3 antibodies against the human leucocyte antigen CD52. Immunology 93, 595–600.Veerhuis, R., Van Es, L.A. and Daha, M.R. (1985). In vivo degradation of rat C1q induced by intravenous injection of soluble IgG aggregates. Immunology 54, 801-810.Wing, M.G., Seilly, D. J., Bridgman, D.J. and Harrison, R.A. (1993). Rapid isolation and biochemical characterization of rat C1 and C1q. Molecular Immunology 30, 433-440... Read More | Inquire | Inquire |