These are supported by a wide range of ELISA reagents, assay and kits providing relative profiling or absolute quantitation of proteins across a wide range of study areas.
When coupled with the breadth of existing global expertise in immunoassays, it is easy to see why ELISA still remains the methodology of choice by those labs tasked with analyte detection.
The preferred manufacturer of these ELISA washers was Bio Tek (Figure 5).
The majority (51%) of survey respondents were using the 96-well microplate as their primary assay format for ELISA assays today (2012).
Among those responding no interest was found for blood banking, stem cells, environmental or quality control as the main (primary) applications of ELISA assays (Figure 1).
Of the sample source analysed by ELISA most respondents (67%) were analysing serum.Other automated instrumentation is designed to support the creation/coating of ELISA assay plates or contains cartridge-based delivery of assay reagents to reduce set-up time.New developments in automated plate washing and the minimisation of wash steps are also impacting conventional ELISAs.erhaps most significantly are the big increases in assay sensitivity and the widening of the dynamic range accomplished by using alternatives to the traditional colorimetric detection, or by the use of new platform technologies.Several of the new offerings utilise bead or particle-based components some of which are advantageous for multiplexing.The majority (57%) of survey respondents do not see the need for commercially available ELISA kits to be miniaturised, indicating that the standard volume 96-well format meets their current needs.Of those seeking miniaturisation, a standard volume 384-well format (14% wanting) or a microchannel 96-well format (eg Siloam Biosciences Optimiser plates) were preferred (12% wanting) (Figure 7).The enzyme/detection chemistries most used by survey respondents today (2012) in their ELISA assays was HRP (horseradish peroxidase)-colorimetric (88% using); followed by AP (alkaline phosphatase)- colorimetric (31% using) and then HRPchemiluminescent (29% using).In contrast, only 13% were using electrochemiluminescent detection (Mesoscale Discovery) and 8% using Alpha Screen/Alpha LISA (Perkin Elmer) (Figure 3).For most of this time a heterogeneous, solid-phase microplate-based format utilising multiple washing steps and a colorimetric readouts has predominated.However, numerous variants of this format have been developed over the years (eg indirect, sandwich, competitive, etc) and a multitude of detection chemistries have been applied to ELISA, including chemiluminescence and various fluorescent readouts.
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