| Description | Unlike most TGF-β family members, GDF-3 and GDF-9 are not disulfide-linked dimers. GDF-3 is expressed in adult bone marrow, spleen, thymus, and adipose tissue. The expression of GDF-3 is upregulated in high-fat-fed wild-type FABP4/aP2 null mice and was associated with obesity, but not with the Unlike most TGF-β family members, GDF-3 and GDF-9 are not disulfide-linked dimers. GDF-3 is expressed in adult bone marrow, spleen, thymus, and adipose tissue. The expression of GDF-3 is upregulated in high-fat-fed wild-type FABP4/aP2 null mice and was associated with obesity, but not with the related hyperglycemia/hyperinsulinemia which characterizes Type 2 diabetes... Read More | Amine-Reactive probe which passively diffuse into cells and it is nonfluorescent until the acetate groups are cleaved by intracellular esterases to yield the highly fluorescent, amine-reactive fluorophore. Upon reaction with amine-containing residues of intracellular proteins, these probes form dye Amine-Reactive probe which passively diffuse into cells and it is nonfluorescent until the acetate groups are cleaved by intracellular esterases to yield the highly fluorescent, amine-reactive fluorophore. Upon reaction with amine-containing residues of intracellular proteins, these probes form dye protein adducts that are well retained in cells as they move and divide during embryonic development.A Non-fluorescent cell permeant amine-reactive probe for long term tracing of cell... Read More | Biochemical Test:SDS-PAGE (purity > 80%); Western blot with patient sample.Calculated Isoelectric Point:pH 6.10 | Protein Purity>90 % by SDS PAGEExtinction CoeffA280 nm = 0.725 at 1.0 mg/mL for pure C1s-C1INH ComplexMolecular Weight196,000 Da (1 chain)General DescriptionThe product C1s-C1INH Complex is made by interacting purified protease inhibitor C1-INH with purified C1s enzyme followed by purification. Protein Purity>90 % by SDS PAGEExtinction CoeffA280 nm = 0.725 at 1.0 mg/mL for pure C1s-C1INH ComplexMolecular Weight196,000 Da (1 chain)General DescriptionThe product C1s-C1INH Complex is made by interacting purified protease inhibitor C1-INH with purified C1s enzyme followed by purification. The protease inhibitor C1-INH prevents the spontaneous activation of complement and limits consumption of C2 and C4 by rapidly inactivating C1r, C1s and MASP2. It is the only plasma serine protease inhibitor (Serpin) capable of interacting with and inhibiting activated C1. C1-INH interacts with the catalytic sites of both C1r and C1s. The interaction with activated C1r and C1s is covalent resulting in complexes which are stable to SDS. C1s and C1r enzymes, however, are irreversibly inactivated by binding to C1-INH. C1s-C1INH is a very stable complex that remains intact even when subjected to freeze/thaw cycles with almost no loss of the complex form.Physical Characteristics & StructureThe C1s enzyme-C1INH complex is composed of two disulfide linked chains from C1s enzyme (A chain 58,000 Da and B chain 28,000 Da) and one covalently linked chain from C1-INH (75,000 Da).SDS-PAGE analysis of the C1s-C1INH complex shows a single band of about 161,000 Da under nonreducing conditions. Under reducing conditions, the C1s-C1INH complex exhibits two bands: A 58,000 Da band corresponding to the A chain of C1s enzyme and a second 103,000 Da band resulting from C1INH (75,000 Da) covalently bond to the B chain (28,000 Da) of C1s enzyme.RegulationActivated C1s is controlled by C1-INH. C1s enzyme and C1-INH form a covalent complex that is resistant to separation on SDS gels. During complement activation C1 complex is rapidly activated by binding to immune complexes. The resulting activated C1s and C1r are rapidly inactivated by interaction with C1-INH (Ziccardi, R.J. (1982)). Binding to immune complexes is fast (10-20 sec) and activation of the bound C1 complex takes several minutes, but C1-INH has also been shown to be fast and no active C1r or C1s remain 4 min after addition of immune complexes to plasma (Ross, G.D. (1986); Ziccardi,R.J. (1981)). The binding of C1-INH to activated C1 releases both C1r and C1s from the complex leaving C1q bound to the immune complex. The released complexes contain four molecules: C1-INH-C1r-C1s-C1-INH. The reaction of C1 esterase inhibitor with activated C1 is very fast with the estimated half-life of C1r and C1s being approximately 15 seconds in serum. In fact, at serum concentrations of C1- INH little or no additional C4 or C2 activation occurs 3 min after immune complexes are added because all the C1r and C1s molecules have been inactivated and removed from the C1q which remains bound to the immune complex (Ross, G.D. (1986); Morley, B.J. and Walport, M.J. (2000); Rother, K., et al. (1998); Ziccardi, R.J. (1982a and 1982b); Morgan, B.P. (1990)). The interaction of purified C1s enzyme and C1-INH is slower.FunctionSee General Description and Regulation above.ApplicationsC1s-C1INH complex can be used in studies designed for developing and identifying inhibitors of C1s-C1INH complex formation and thus lead to the possible development of therapeutics for inhibiting complement activation via the classical pathway.GeneticsThe EMBL/Genbank cDNA accession number for C1s is J04080. The gene for C1s is located on chromosome 12p13. The EMBL/Genbank cDNA accession numbers for C1-INH are M13656 and X54486 (human) and Y10386 (mouse). The gene for C1-INH is located on chromosome 11p11.2-13. DeficienciesC1s deficient patients are prone to systemic lupus erythematosus (SLE) and recurrent pyogenic infections (Rother, K., et al. (1998)). They lack classical pathway function. The genetic disorder hereditary angioedema (HAE) is caused by a partial deficiency of C1-INH. Patients with HAE have low functional C1-INH levels in blood and have recurrent episodes of systemic or localized edema.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.ReferencesZiccardi, RJ. (1982) A new role for C-1-inhibitor in homeostasis: control of activation of the first component of human complement. J. Immunol. 128:2505-2508.Ross, G.D. (1986) Immunobiology of the Complement System. (ISBN 0-12-5976402) Academic Press, Orlando.Ziccardi, R.J. (1981) Activation of the early components of the classical complement pathway under physiologic conditions. J. Immunol. 126:1769-1773.Morley, B.J. and Walport, M.J. (2000) The Complement Facts Book. (ISBN 0127333606) Academic Press, London.Rother, K., Till, G.O., and Hӓnsch, G.M. (1998) The Complement System. (ISBN 3-540- 61894-5) Springer-Verlag, Heidelberg.Ziccardi, R.J. (1982a) Spontaneous activation of the first component of human complement (C1) by an intramolecular autocatalytic mechanism. J. Immunol. 128:2500- 2504.Ziccardi, RJ. (1982b) A new role for C-1-inhibitor in homeostasis: control of activation of the first component of human complement. J. Immunol. 128:2505-2508. Morgan, B.P. (1990) Complement Clinical Aspects and Relevance to Disease. (ISBN 0- 12-506955-3) Academic Press, London... Read More | 1、Product attributeShelf life: 24 monthsReaction time：long (up to 45 minutes) at 20-37°CLot-to-lot variation：<10%Boiling point : 100℃pH-Value (at 20 °C): 9.0-9.8 Density (20℃) : 1.0302 g/cm³Water solubility: easily solubleAppearance: colourless to 1、Product attributeShelf life: 24 monthsReaction time：long (up to 45 minutes) at 20-37°CLot-to-lot variation：<10%Boiling point : 100℃pH-Value (at 20 °C): 9.0-9.8 Density (20℃) : 1.0302 g/cm³Water solubility: easily solubleAppearance: colourless to light yellow liquidOdour: odourlessIncubation temperature: 20-37 °CLight sensitiveHeat sensitive 2、Requirements for storage rooms and vessels1.Keep container tightly closed.2.Keep cool. protected from light3. Do not store together with: Oxidizing agent. 4. Contaminated or leaked out substrate solution from damaged bottles should not be used anymore and has to be destroyed.5. Use isolated containers with some cool bags for transport.6. Spontaneous decay will increase the background. If stored at room temperature, the velocity of the decay will increase. Thus, both storage and transport at room temperature should be avoided. Nevertheless, the activity of the solution is not affected by storage at room temperature. The solution still works beyond the expiry date, but some applications, especially those including visual evaluation, may be hampered by increased background. 3、Effective Components and Principle of FunctionIn different buffer solutions (pH = 9.5), with supplementation if required, the effective componentpara nitrophenyl phosphate (pNPP) is dissolved. Alkaline Phosphatase transfers the phosphate residue to an acceptor. Under alkaline conditions a yellow colour occurs, resulting from the formed nitrophenol. 4、Biosafety informationThis mixture is not classified as hazardous in accordance with Regulation (EC) No 1272/2008； 5、Advantage1. Signal yield comparable to competitor ready-to-use solutions2. Broad measurement range3. Very low background signals4. Very low blank drift during long-term storage (<0.15 AU within 24 months)5. High colour stability after reaction stop with this product and other commonly used stopping solutions 6、Instruction for usageFor bottling consider the following instructions:• Work in a dust free and darkened room.• Keep the solution as cool as possible.• Avoid contact of the solutions with any metal parts• Clean all instruments and vessels very extensively.• Wear powder-free gloves during bottling.• Close the bottles immediately to minimize the influence of light and dust.• Use clean bottles that are impermeable to light made from HDPE or PP. 7、 General Instructions for the Use in Blotting Systems Only qualified laboratory staff, who are familiar with the basics of immunological methods, are allowed to use these solutions.The substrate solutions can be used in qualitative and quantitative ELISA procedures.When using 96-well microtiter plates, adding 100 µL of substrate per well after incubation and washing is recommended. After substrate incubation the reaction can be stopped and the photometric measurement can be carried out. Using higher incubation temperatures (37° C) may shorten the incubation time. The reaction can be stopped by using the special developed solution stop. The use of other commercially available stop solutions cannot safely exclude a further increase of the signal. Addition of a stopping solution does not change the general shape of the spectrum. The unstopped and the stopped solution should be measured at 405 nm and the background correction: should be measured at 620 nm... Read More |