| 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 | 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 | Inquire | Inquire |