Serum, plasma, cell culture supernatant and other biological samples
Species Reactivity
Monkey
Detection Method
sELISA
Intended Use
In vitro quantitative determination of C5b-9 concentrations in serum, plasma, cell culture supernatant and other biological samples.
Contents of Kit
The sealed kit can be stored at 2-8°C. The storage condition for opened kit is specified in the table below: 1. ELISA Microplate(Dismountable), 8×12. Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C. Stored for 12 month at -20°C 2. Lyophilized Standard, 2vial. Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C. Stored for 12 month at -20°C. 3. Biotin-labeled Antibody(Concentrated, 100×), 120μl. 2-8°C (Avoid Direct Light) 4. HRP-Streptavidin Conjugate(SABC, 100×), 120μl. 2-8°C (Avoid Direct Light) 5. TMB Substrate, 10ml. 2-8°C (Avoid Direct Light) 6. Sample Dilution Buffer, 20ml. 2-8°C 7. Antibody Dilution Buffer, 10ml. 2-8°C 8. SABC Dilution Buffer, 10ml. 2-8°C 9. Stop Solution, 10ml. 2-8°C 10. Wash Buffer(25×), 30ml. 2-8°C 11. Plate Sealer, 5 pieces 12. Product Description, 1 copy Note: The liquid reagent bottle contains slightly more reagent than indicated on the label. Please use pipette accurately measure and do proportional dilution.
Storage
2-8°C (for sealed box), please do not freeze! See kit label for expiry date
Performance Characteristics
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
Precision
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on same plate. Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Detection Range
0.313-20ng/ml
Sensitivity
0.188ng/ml
General Description
C5b-9 Activation of the complement system leads to the formation of C5b-9 terminal complex, and while C5b-9 can promote cell lysis, the sublytic assembly of C5b-9 on plasma membranes causes an opposite result and induces cell cycle activation and survival. C5b-9 can rescue oligodendrocytes from FAS-mediated apoptosis by regulating caspase-8 processing via PI 3-K signaling. C5b-9 may play a pro-inflammatory role in the acute phase of multiple sclerosis, but may also be neuroprotective during the chronic phase of the disease.
Standard Curve
This product has been tested by Quality Control Department and meets performance specifications mentioned in the manual. (The humidity in the laboratory is 20%-60%, and the temperature is 18°C -25°C. TMB was balanced to 37°C before color development, and incubated at 37°C for 15 minutes in the dark after adding the enzyme label plate holes.) The following assay data are provided for reference, since experimental environment and operation are different. The establishment of standard curve depends on your own assay.
Citations
Publication ()
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Background
The complement system contains a wide variety of different proteins that interact in a highly organized manner in an intricate crosstalk with innate and adaptive immune pathways. Complement components are arranged in a cascade, ready to amplify and respond rapidly to various microenvironmental insults or cues. Among the many complement components, C3 and C5 are important in the complement cascade. In general, the cascade can start with three different pathways: the classical pathway (CP), the alternative pathway (AP) and the lectin pathway (LP). The classical pathway begins with the binding of the C1q fragment of complement factor C1 to antigen-antibody complexes. The lectin pathway begins with the binding of mannose-conjugated lectin to polysaccharide or glycoprotein moieties on the surface of foreign cells or injured host cells. In contrast, AP is characterized by constitutive low-level activation of C3. This activation, known as the "C3 tickover," is mediated by the spontaneous hydrolysis of C3, which leads to the generation of C3H20 and the formation of the active C3 converting enzyme, C3H20Bb. The C3H20Bb converting enzyme initiates the AP.
Figure 1. Overview of the complement system (Source: Kim BJ, et al. 2021)
The C5 converting enzyme C3bBbC3b lysis of C5, releasing C5b and C5a. C5b initiates the formation of the C5b-9 membrane attack complex (MAC), whereas C5a exerts multiple pro-inflammatory and immunomodulatory functions. C5b-9 is the end product of the classical, lectin and alternative complement pathways and is thought to play an important role in the pathogenesis of various renal diseases, causing cellular damage and tissue inflammation leading to sclerosis and fibrosis. MAC is a key downstream effector of the complement cascade and has the ability to lysis target cells by forming pores in the cell membrane or to activate intracellular signaling pathways at sublethal doses. C5b-9 formation can be incompletely arrested without anchoring to the membrane, in which case it circulates as a soluble complex with vitronectin or clusterin, termed sC5b-9.
Both C5b-9 and sC5b-9 promote inflammation and thrombosis. Blood and urine sC5b-9 levels are elevated in a variety of kidney diseases and correlate with disease activity and severity. In all renal diseases, deposits of C5b-9 are predominantly found in areas of glomerulosclerosis, tubulointerstitial injury, vascular hyalinization and sclerosis. This finding was most pronounced in hypertensive and diabetic nephropathy, interstitial nephritis and acute tubular necrosis. In clinical applications, sC5b-9 is easily formed in vitro and has a short half-life, making it inconvenient to use for clinical testing. The investigators believe that the deposition of C5b-9 in the kidney better reflects its formation in the pathogenesis of kidney disease.
Alternative Names
Monkey membrane attack complex ELISA Kit Monkey MAC ELISA Kit
References
1. Kim BJ, et al. Targeting complement components C3 and C5 for the retina: Key concepts and lingering questions. Prog Retin Eye Res. 2021 Jul;83:100936.
2. Koopman JJE, et al. Deposition of the Membrane Attack Complex in Healthy and Diseased Human Kidneys. Front Immunol. 2021 Feb 11;11:599974.
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References
The Dimeric Form of 1,3-Diaminoisoquinoline Derivative Rescued the Mis-splicing ofAtp2a1andClcn1Genes in Myotonic Dystrophy Type 1 Mouse Model
Expanded CUG repeat RNA in the dystrophia myotonia protein kinase (DMPK) gene causes myotonic dystrophy type 1 (DM1) and sequesters RNA processing proteins, such as the splicing factor muscleblind-like 1 protein (MBNL1). Sequestration of splicing factors results in the mis-splicing of some pre-mRNAs. Small molecules that rescue the mis-splicing in the DM1 cells have drawn attention as potential drugs to treat DM1. Herein we report a new moleculeJM642consisted of two 1,3-diaminoisoquinoline chromophores having an auxiliary aromatic unit at the C5 position.JM642alternates the splicing pattern of the pre-mRNA of theLdb3gene in the DM1 cell model andClcn1andAtp2a1genes in the DM1 mouse model.In vitrobinding analysis by surface plasmon resonance (SPR) assay to the r(CUG) repeat and disruption of ribonuclear foci in the DM1 cell model suggested the binding ofJM642to the expanded r(CUG) repeat in vivo, eventually rescue the mis-splicing.
Synthesis and Structures of Polyphenylphenanthrenes
CHEMISTRY-A EUROPEAN JOURNAL
Authors: Xiao, Yonglong; Mague, Joel T.; Donahue, James P.; Wilson, Laura J.; Kraml, Christina M.; Pascal, Robert A., Jr.
1,2,3,4,5,6,7,8-Octaphenylphenanthrene (4) and decaphenylphenanthrene (5) were prepared by very short syntheses (two or three steps) from tetraphenylfuran and polybrominated benzene derivatives. The X-ray structures of compounds 4and5show them to be quite crowded, with the phenanthrene cores twisted by about 40 degrees due to the clash of the C4 and C5 phenyl groups. Compound 4was resolved by chromatography on a chiral support, and its free energy of activation for racemization was determined to be 24.6 kcal mol(-1)at 40 degrees C. Computational studies indicate that compound 5has a racemization barrier approximately 6 kcal mol(-1)lower than4, and thus5would not be configurationally stable at room temperature.
COA
Publication 
Figure 1. Overview of the complement system
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