| Description | Absorbance: A412 nm = 0.87 at 1/50 dilution in deionized waterGeneral DescriptionSheep red blood cells which have been washed free of sheep complement proteins, but were not coated with hemolysin antibodies. These cells have traditionally been used as negative controls for assays of the human Absorbance: A412 nm = 0.87 at 1/50 dilution in deionized waterGeneral DescriptionSheep red blood cells which have been washed free of sheep complement proteins, but were not coated with hemolysin antibodies. These cells have traditionally been used as negative controls for assays of the human classical and alternative complement pathways in serum samples (Morgan, B.P. (2000); Dodds, A.W. and Sim, R.B. (1997)). Most human serum samples have small amounts of natural antibodies (usually IgG) to sheep antigens and at high concentrations of serum will agglutinate Es and activate the classical pathway leading to lysis. This rarely occurs at dilutions used in the CH50 titration assay where serum is diluted more than 1/100. Es are supplied at assay-ready concentrations in the traditional buffer used in CH50 assays (GVB⁺⁺). They can usually be used for 2 weeks after preparation. They are shipped cold, but are not harmed by extended periods at room temperature (note that they circulate 60-90 days a 37℃ in vivo). They should be washed once before each use (5 min at 500 to 1000 x g at 4℃) and resuspended in GVB++ to reduce background. This procedure may also be used to concentrate the cells.Physical CharacteristicsEs are natural uncoated erythrocytes that do not activate complement under typical assays conditions. They do not have IgG or IgM on their surface, however, there are natural antibodies in most animal serum. In the most sensitive assays it has been estimated that as few as 10 IgM/EA can cause lysis while this requires approximately 1000 IgG/EA (Ross, G.D. (1986)). This is primarily because in IgM there are five closely spaced Fc domains in fixed positions on each molecule. The spacing of IgG molecules is more random and it is relatively rare to find two or more IgG in the correct positions on the surface of a cell.ApplicationsEs cells are primarily used as controls for CH50 assays and for specialized tests requiring uncoated erythrocytes. Natural antibodies present in human blood to animal antigens may cause agglutination of the cells and lysis if the serum is used at high concentrations, however, this does not interfere with the CH50 titers because EA used in these assays already carry high levels of IgM antibodies and serum is diluted 100-fold or more. IgG antibodies are 100-fold less active than IgM and most of the cross-reactive antibodies in human blood are IgG.RegulationSheep erythrocytes (Es) are used for human complement assays partly for convenience, but also because they lack membrane-bound regulators of human complement. No significant level of functional DAF, CD59 or CR1 exists on Es for human complement. Thus, Es are useful for their lack of membrane regulatory activities... 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 | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： This protein is a cell adhesion molecule involved in neuron-neuron adhesion, neurite fasciculation, outgrowth of neurites, etc | Purity:>90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Implicated in the control of cell proliferation and cellular aging. May also act as a chaperone | Purity>98% by SDS-PAGE and HPLC analyses.FunctionChemoattractant for blood monocytes, memory T-helper cells and eosinophils. Causes the release of histamine from basophils and activates eosinophils. Binds to CCR1, CCR3, CCR4 and CCR5. One of the major HIV-suppressive factors produced by CD8+ T-Purity>98% by SDS-PAGE and HPLC analyses.FunctionChemoattractant for blood monocytes, memory T-helper cells and eosinophils. Causes the release of histamine from basophils and activates eosinophils. Binds to CCR1, CCR3, CCR4 and CCR5. One of the major HIV-suppressive factors produced by CD8+ T-cells. Recombinant RANTES protein induces a dose-dependent inhibition of different strains of HIV-1, HIV-2, and simian immunodeficiency virus (SIV). The processed form RANTES(3-68) acts as a natural chemotaxis inhibitor and is a more potent inhibitor of HIV-1-infection. The second processed form RANTES(4-68) exhibits reduced chemotactic and HIV-suppressive activity compared with RANTES(1-68) and RANTES(3-68) and is generated by an unidentified enzyme associated with monocytes and neutrophils... Read More |