Preparing Fluorescence Assays While Avoiding Errors

Standard Preparation

  1. The assay type plus both the excitation and emission wavelengths should be determined. This can be based on the sample for analysis or the desired results.
     
  2. The highest gain and sensitivity should be chosen to allow for a maximum line arrange.
     
  3. A uniform volume of each reagent should be pipetted into the selected wells of a 96 well plate. Volumes must be identical as any well-to-well variations may cause significant error. The minimum required volume of each reagent should be used.
     
  4. Replicates of a standard curve should be prepared and used as a reference for concentration determination.
     
  5. Once all the reagents have been added, the wells should be thoroughly mixed through small pipette bursts. The formation of bubbles should be avoided as these may cause a light scattering effect when reading the plate. A needle or a small pipette tip can be used to pop any bubbles.
     
  6. The plate should be centrifuged through the use of the OHAUS 5718R Multi-Pro Refrigerated Centrifuge (30314815) with the Microplate Rotor Attachment (30314824).
     
  7. Bright samples should be segregated through the use of empty wells to avoid error.
    The outer wells should be left empty as a buffer from plate bleed over.  
     
  8. Once enough time has elapsed for the assay to complete, a fluorescence plate reader should be used to obtain the final results.
     
  9. Once read, the plate should be disposed of as exposure to fluorescent light can cause photo bleaching of the sample.

OHAUS Products Used Within this Procedure

5718R Multi-Pro Refrigerated Centrifuge

5718R Multi-Pro Refrigerated Centrifuge

Microplate Rotor Attachment

Microplate Rotor Attachment

About OHAUS Corporation

OHAUS Corporation manufactures an extensive line of weighing products, laboratory equipment and analytical instruments that meet the weighing and measurement needs of virtually every industry.

They are a global leader in the laboratory, industrial and education channels, as well as a host of specialty markets, including the food preparation, pharmacy and jewelry industries. An ISO 9001:2008 manufacturer, OHAUS products are precise, reliable and affordable, and backed by industry-leading customer support.


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Last updated: Nov 5, 2019 at 7:41 AM

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