Description
Factor V Paired Antibody Set
Affinity’s Factor V Paired Antibody Set consists of matched capture and detecting antibodies that have been titrated and optimized for use in sandwich style ELISA assays. The product as provided contains sufficient capture and detecting antibodies for five full 96-well microplates and contains a detailed protocol sheet containing directions for use, recipes for solutions and sources for additional materials required. This Factor V Paired Antibody Set is intended to facilitate the end user in establishing an “in-house” immunoassay for research purposes only and must not be used for diagnostic applications. Assay validation is the responsibility of the end user.
Product Code: FV-EIA
Supplied Materials:
- Capture Antibody (FV-EIA-C): One yellow-capped vial containing 0.5 ml of polyclonal affinity purified anti-Factor V antibody for coating plates.
- Detecting Antibody (FV-EIA-D): One red-capped tube containing 0.5 ml of peroxidase conjugated affinity-purified polyclonal anti-Factor V antibody for detection of captured Factor V.
Related Products: VisuLize Buffer Pak
Species Cross Reactivity: View Chart
Product Datasheet: Factor V (F5 FV) Antigen Matched Pair Antibody Set for ELISA - FV-EIA
Description of Factor V
Factor V (formerly referred to as accelerator globulin and labile factor) is a large glycoprotein (320 kDa) that is produced in the liver. The gene that encodes factor V (FV) is located on chromosome 1. A congenital deficiency of FV is a hemorrhagic disorder inherited as an autosomal recessive disease. The concentration of FV in plasma is typically 10 μg/mL. FV is a pro-cofactor that is activated through limited proteolysis by thrombin, or by activated factor X in the presence of phospholipid surface. Other physiologic activators of FV include plasmin, neutrophil elastase and platelet calpain. The activated cofactor (FVa) is an essential component of the prothrombin activator complex, which consists of FVa, activated factor X, calcium and anionic phospholipid surface. The intact prothrombinase complex activates prothrombin to thrombin at a rate 300,000-fold greater than activated factor X alone. In a positive feedback loop, the thrombin generated accelerates its own generation by activating more FV to FVa. Thrombin also acts to down-regulate FVa indirectly by activating Protein C, which inactivates FVa cofactor activity1-3.
References and Reviews
- Kane WH, Davie EW; Blood Coagulation Factors V and VIII: Structural and functional similarities and their relationship to hemorrhagic and thrombotic disorders. Blood 71:539, 1988.
- Hoyer, LW, Wyshock EG, Colman RW, in Hemostasis and Thrombosis, 3rd Edition, eds. RW Colman, J Hirsh, VJ Marder and EW Salzman, pp. 109-133, J.B. Lippincott Co., Philadelphia, 1994.
- Nesheim ME, Katzmann JA, Tracy PB, Mann KG; in Methods in Enzymology 80:249, 1980.
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比如你用anti-MHC-II,mouse IgG1-FITC,那同型就用非特异性的mouse IgG1-FITC,同样浓度
阴性对照(Isotype Control):非特异荧光的强弱取决于抗体浓度、单克隆荧光抗体特异性和纯度,应与试验管抗体相对应。在多色分析时,同型对照应与其它抗体同时使用,以避免补偿造成的误差
血小板体外活化试验:使用正常人活化标本作为阳性质控;使用正常人未活化标本作为阴性质控
血小板自身抗体检测:使用含有已知血小板抗体的血清与血小板孵育,作为阳性质控;使用不含血小板抗体的血清与血小板孵育,作为阴性质控
血小板表面抗原缺失:如巨血小板症血小板表面CD42a/CD42b缺失,血小板无力症血小板表面gpIIb/IIIa,即CD41/CD61缺失或异常。使用正常人标本做阳性对照,抗体的同型对照做阴性对照
选用和特异性抗体同种亚型的,同样荧光素标记的抗体,采用同样的染色步骤染同样浓度的细胞。然后上机检测。如果细胞有非特异性的结合,那同型对照的荧光强度可能会比未染色的阴性对照要高一些。
同型对照只用于那种阳性和阴性之间没有明确分解的情况。如果阳性细胞群非常清楚,是不需要同型对照的。另外,在多色同染的情况下,也不采用同型对照,而是采用FMO(fluorensence minus one)对照。
另外,对于同型对照的使用,也有很多反对意见。就算是同型抗体,也可能会有不同的亲和力,并不一定就能真实反应非特异性的结合。
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