Bradykinin ELISA试剂盒| Abcam中文官网

来自 : 发布时间：2024-05-14

产品概述 Bradykinin ELISA kit is a competitive Enzyme-Linked Immunosorbent Assay designed for the accurate quantitative measurement of Bradykinin in plasma, serum and urine.A goat anti-Rabbit IgG antibody has been precoated onto 96-well plates. Standards or test samples are added to the wells, along with a solution of Bradykinin conjugated to biotin, followed by a solution of polyclonal antibody to Bradykinin. The plate is washed to remove unbound reagents. A solution of streptavidin-HRP conjugate is then added. After further incubation the excess reagents are washed away and TMB substrate is added, which is catalyzed by HRP to generate a yellow color. A stop solution changes this color from yellow to blue, and the intensity of this blue coloration is inversely proportional to the amount of Bradykinin captured in the plate. 2 vialsGoat anti-rabbit IgG Microplate (12 x 8 wells) 1 unitHRP- Streptavidin Conjugate 1 x 12.5µgPlate Sealer 2 unitsStop Solution 2 1 x 10mlTMB Substrate 2 x 10ml (1) Kininogens are inhibitors of thiol proteases; (2) HMW-kininogen plays an important role in blood coagulation by helping to position optimally prekallikrein and factor XI next to factor XII; (3) HMW-kininogen inhibits the thrombin- and plasmin-induced aggregation of thrombocytes; (4) the active peptide bradykinin that is released from HMW-kininogen shows a variety of physiological effects: (4A) influence in smooth muscle contraction, (4B) induction of hypotension, (4C) natriuresis and diuresis, (4D) decrease in blood glucose level, (4E) it is a mediator of inflammation and causes (4E1) increase in vascular permeability, (4E2) stimulation of nociceptors (4E3) release of other mediators of inflammation (e.g. prostaglandins), (4F) it has a cardioprotective effect (directly via bradykinin action, indirectly via endothelium-derived relaxing factor action); (5) LMW-kininogen inhibits the aggregation of thrombocytes; (6) LMW-kininogen is in contrast to HMW-kininogen not involved in blood clotting. 组织特异性 Secreted in plasma. T-kinin is detected in malignant ovarian, colon and breast carcinomas, but not in benign tumors. 疾病相关 Defects in KNG1 are the cause of high molecular weight kininogen deficiency (HMWK deficiency) [MIM:228960]. HMWK deficiency is an autosomal recessive coagulation defect. Patients with HWMK deficiency do not have a hemorrhagic tendency, but they exhibit abnormal surface-mediated activation of fibrinolysis. 翻译后修饰 Bradykinin is released from kininogen by plasma kallikrein.Hydroxylation of Pro-383 occurs prior to the release of bradykinin.Phosphorylation sites are present in the extracelllular medium.N- and O-glycosylated. O-glycosylated with core 1 or possibly core 8 glycans. 发表研究结果有使用 ab136936？请让我们知道，以便我们可以引用本数据表中的参考文章。 ab136936 被引用在 3 文献中. Matsuura H et al. C1 Esterase Inhibitor Activity Is Reduced in the Acute Phase Following Burn Injury: A Prospective Observational Study. J Burn Care Res 40:893-899 (2019).PubMed: 31250897 Ito M et al. Prostanoid-dependent spontaneous pain and PAR2-dependent mechanical allodynia following oral mucosal trauma: involvement of TRPV1, TRPA1 and TRPV4. Mol Pain 13:1744806917704138 (2017).PubMed: 28381109 Vieira ML et al. Modulation of Hemostatic and Inflammatory Responses by Leptospira Spp. PLoS Negl Trop Dis 10:e0004713 (2016).PubMed: 27167223 Objective:We used this kit to test bradykinin concentrations in serum and EDTA plasma samples from human and rhesus macaque. No samples were Ethanol treated. We also performed a spike recovery on diluent and both rhesus serum and plasmaEase of Use:We were confused on some issues regarding the controls and methods of the procedures. There have been several versions of the SOP in recent times, and we found that Version 3, which came with the kit, had already been supplanted by Version 4 and 5 within months. The directions and recommendations varied. The EDTA plasma preparation recommendations are confusing: \"Mix blood in a ratio of 1:4 with ice cold ethanol.\" Should this be 1 part with 4 parts, or 1 part of 4 parts? \"Dry down sample...\" What? How? How can it be prevented from degrading during this \"dry-down\"? The different treatments for the control wells add points of common error, and the explanation on how to interpret these controls is not readily understood, particularly to those without experience with competitive ELISAs.Interpretation: This kit does detect Rhesus bradykininWe were able to determine that decreasing OD (competitive ELISA) correlated to increasing concentrations of Bradykinin in human and rhesus samples in a similar fashion.Results: EDTA plasma:For n=2 rhesus animals, extrapolated values were the same for dilutions of 1:16, 1:64, and 1:256 n=1 rhesus animal extrapolated values varied at all dilutions. In the corresponding human sample, results for 1:16 and 1:64 corresponded.Results: Serum:There seemed to be no correlation among the tested dilutions for extrapolated values in human or rhesus. We believe this relates to dilution-dependent ex vivo generation during the assay.Spike Recover:There was virtually no relationship between the amount spiked and the amount measured in the rhesus serum or plasma (notice the logarithmic scale for that test). We did not repeat this assay with human serum or plasma.Values in these graphs are average +/- 1 standard deviationab136936: ABTRIAL-TVB33 value: $573 It is preferable to perform the extraction/handling as it is laid out in the kit protocol as this will help to remove interference as well as other factors that can contribute to the instability of the sample. Read More A pesar de que hay is optimizado las diluciones para vuestras muestras, las diluciones que hab is usado est n por debajo del m nimo recomendado para evitar el efecto matriz. Os sugiero una diluci n m nima de 1/16 para plasma y orina. De los resultados enviados observamos un alto coeficiente de variaci n entre duplicados. Esto suele deberse a problemas ocurridos durante el pipeteo o a algo que se haya a adido al tamp n de diluci n que pueda estar interfiriendo en las medidas. La curva de est ndares es pr cticamente plana, pero lo m s preocupante es que varios de los puntos de la curva superan en absorbancia al valor de B0, lo cual es imposible en la pr ctica. B0 es el punto en el que el 100% del anticuerpo esta unido al est ndar, y por tanto da el valor m s alto de absorbancia en muestras y est ndares, ya que no hay Bradykinina para competir por el anticuerpo.El hecho de que este valor sea m s bajo que otros puntos de la recta implica que algo ha ido mal durante el ensayo o la configuraci n de la placa durante el mismo.Est is usando alg n otro diluyente aparte del Assay Buffer del kit para diluir el est ndar? Una vez preparada los est ndares para la curva, se tardaron m s de 30 minutos en tomar las medidas de los mismos? En caso contrario el est ndar pudo haberse degradado.Se sigui el esquema de concentraciones de est ndares indicado en el protocolo o se prepararon diluciones diferentes?Estaban todos los reactivos a temperatura ambiente antes de usarlos, habi ndolos sacado durante al menos 30 minutos antes de comenzar el ensayo? En caso contrario la eficiencia de uni n del anticuerpo puede disminuir.Se elimin completamente el buffer de lavado de los pocillos antes de a adir el reactivo? Es importante asegurarse que no quedan restos de buffer en los pocillos, pero teniendo cuidado de no secar la placa (si se invierte la placa y golpea para eliminar el exceso de buffer de lavado, debe hacerse r pido y con un par de golpecitos simplemente).Os animo a repetir la curva de est ndares, si aun es posible, y probar con varias diluciones de una muestra representativa para comprobar si los valores siguen sin ser los esperados. Aunque pueda parecer evidente tambi n os recomiendo comprobar que la configuraci n de la placa es la adecuada y que el lector est configurado de forma apropiada para leer a la longitud de onda indicada. Read MorePlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com