CRP canine ELISA
產(chǎn)品名稱：CRP canine ELISA
產(chǎn) 地：Demeditec
產(chǎn)品貨號：DE931CRP01D
產(chǎn)品規(guī)格：96 Tests
產(chǎn)品說明：
1. Angela Veiga, PhD
Faculdade de Veterinária, Departmento de Patologia Clínica
Universidade Federal do Rio Grande do Sul (UFRGS), Brazil:
C-reactive protein (CRP) is a positive acute phase molecule secreted by the liver in response to internal and external injury. CRP binds to membrane receptors of neutrophils and mononuclear cells, and stimulates the release of proinflammatory cytokines. In dogs, increasing CRP levels were associated with a number of inflammatory, infectious, and post-traumatic conditions, making it a good marker of tissue damage and healing.
The Canine CRP ELISA Kit from Demeditec Diagnostics contains a 96-well plate coated with purified goat anti-dog CRP-IgG, a vial of concentrated conjugate HRP-labeled goat anti-dog CRP-IgG, a vial of high quality CRP standard, one packet of dehydrated wash buffer, a vial of TMB substrate, and a vial of phosphoric acid as stop solution. The assay is highly specific, sensitive and easy to handle; it is stable for one-year when stored at 2-8°C.
To perform the assay, make sure that all the components have reached room temperature before use. In addition to serum and plasma, a number of specimen types (e.g. urine, saliva) can be used in this assay, making it useful in canine medicine, where sometimes it is difficult to collect certain types of samples. If the choice is to use serum or plasma, make sure that diluted samples (1:500) are used. In my analysis I used diluted serum. To accomplish the measurements, 2 µL of sample is enough. If a specimen other than serum or plasma is used, the manufacturer recommends a dilution of 1:2. A standard curve is made using a series of standards made from the standard concentrate provided. 100 µL of diluted sample is added to each well followed by incubation. The plate is washed and the conjugate is added, followed by incubation and washing steps. The chromogen is then added to start the colour development, which appears blue in positive reactions. Finally, the reaction is discontinued with a stop solution, which turns the blue colour into yellow in positive reactions. The absorbance of the plate is read in a spectrophotometer or plate reader at 450 nm.
2. Rudy M. Ortiz, PhD, Assistant Professor
Department of Physiology, University of California, Merced, United States
Demeditec Diagnostics supplies CRP ELISA kits for the measurement of plasma/serum C-reactive protein (CRP) from a number of different species including human, rat, mouse and dog. While the rat and mouse kits are highly specific, in my experience, the canine kit exhibits some cross-reactivity with other mammalian species (such as seals). The kits come in two detection formats: 1) antibody and 2) C-polysaccharide. I have used both formats to detect rat CRP and have obtained similar results, suggesting that either format is suitable for reliable measurement of rat CRP. Furthermore, using an extraction protocol we developed in our lab, we have also used the rat CRP kits to reliably and repeatedly measure urinary CRP fragments. While the product insert mentions that the kit can be used for the detection of CRP in urine, to the best of our knowledge and capabilities, this was only accomplished following extraction of the CRP fragments in the urine. Nonetheless, our ability to modify the assay to fit our purposes demonstrates the kits flexibility to comply with the needs of the lab, which we found to be very beneficial.
All kits are supplied with all the necessary components/reagents to complete the assay including a 96-well microplate (either antibody or C-polysaccharide coated) and a vial of concentrated standard that needs to be diluted. The kit does not supply the reagents or extraction columns to extract urinary CRP, but these are readily available. The kit insert also does not provide information on a suggested extraction protocol. This needs to be developed separately. For plasma/serum and other media (except urine) samples need to be diluted prior to assay, and new users will need to run some test assays to determine the appropriate dilutions for their needs; however, suggested starting dilutions are provided. While the assay instructions only suggest the use of 4 standards ranging from 1.5 – 40 ug/ml for the canine CRP kit and 6 for the rat CRP ranging from 1.6 – 133 ng/ml, we have improved the reliability and reproducibility of our results by simply serially diluting the highest standard 7 times, going beyond the lowest suggested standard. The limit of detection for the canine kit is 3.6 ng/ml and for the rat kit is 2.5 ng/ml, both of which are very good and should easily accommodate most research needs.
The actual assay procedure is very standard and easy to complete, and the wavelength filter at 450 nm is very common, making the assay very adaptable to virtually any lab that has access to a 96-well plate reader. Because the assay requires multiple washings, the use of an automatic plate washer can reduce the total assay time, but even without the washer an assay can be completed in less than 2 hr. Thus, reliable and accurate results can be obtained in a relatively short period.
We have had great success obtaining accurate and reliable results using their different kits (rat and canine CRP) to satisfy our research purposes and are very comfortable with this product.
上海玉博生物技術(shù)有限公司在為生命科學(xué)領(lǐng)域提供豐富的產(chǎn)品與信息資源方面處于國內(nèi)**地位，公司提供的產(chǎn)品涵蓋了二十多個國家近五十萬種產(chǎn)品，而且產(chǎn)品的數(shù)量與信息在不斷的增長和更新，公司提供的產(chǎn)品能夠使生命科學(xué)工作者加快對生物化學(xué)，分子生物學(xué)，細(xì)胞生物學(xué)以及蛋白質(zhì)組學(xué)研究的認(rèn)知，以及分子診斷和臨床醫(yī)學(xué)領(lǐng)域的應(yīng)用。
