Generation of Two-color Antigen Microarrays for the Simultaneous Detection of IgG and IgM Autoantibodies
J Vis Exp. 2016;(115)
Authors: Chruscinski A, Huang FY, Ulndreaj A, Chua C, Fehlings M, Rao V, Ross HJ, Levy GA
Abstract
Autoantibodies, which are antibodies against self-antigens, are present in many disease states and can serve as markers for disease activity. The levels of autoantibodies to specific antigens are typically detected with the enzyme-linked immunosorbent assay (ELISA) technique. However, screening for multiple autoantibodies with ELISA can be time-consuming and requires a large quantity of patient sample. The antigen microarray technique is an alternative method that can be used to screen for autoantibodies in a multiplex fashion. In this technique, antigens are arrayed onto specially coated microscope slides with a robotic microarrayer. The slides are probed with patient serum samples and subsequently fluorescent-labeled secondary antibodies are added to detect binding of serum autoantibodies to the antigens. The autoantibody reactivities are revealed and quantified by scanning the slides with a scanner that can detect fluorescent signals. Here we describe methods to generate custom antigen microarrays. Our current arrays are printed with 9 solid pins and can include up to 162 antigens spotted in duplicate. The arrays can be easily customized by changing the antigens in the source plate that is used by the microarrayer. We have developed a two-color secondary antibody detection scheme that can distinguish IgG and IgM reactivities on the same slide surface. The detection system has been optimized to study binding of human and murine autoantibodies.
PMID: 27685156 [PubMed - as supplied by publisher]
http://ift.tt/2deWIae
Ιατρική : Τα αισθητικά συστήματα της όρασης,ακοής,αφής,γεύσης και όσφρησης.
Εγγραφή σε:
Σχόλια ανάρτησης (Atom)
Δημοφιλείς αναρτήσεις
-
[PDF] Καρκίνος του στομάχου -- Αλέξανδρος Γ. Σφακιανάκης Αναπαύσεως 5 Άγιος Νικόλαος Λασιθίου 72100 2841026182
-
A Genome‐wide Association Study of Prostate Cancer in Latinos Latinos represent less than 1% of samples analyzed to date in genome‐wide asso...
-
Does CBD Oil Lower Blood Pressure? This article was originally published at SundayScaries." Madeline Taylor POSTED ON January 13, 20...
-
A rare case of an intramedullary metastasis of a myxopapillary ependymoma Category: Unique Case Observations Article type: Case Report Autho...
-
https://sfakianakisorl.blogspot.com/2019/07/a-heavier-weight-at-birth-is-associated.html Alexandros Sfakianakis Anapafseos 5 . Agios Nikolao...
Δεν υπάρχουν σχόλια:
Δημοσίευση σχολίου