1 / 9

Dynatech MR5000

Dynatech MR5000. Using a Microplate Reader for a variety of tasks. Features of the Dynatech MR5000. Microprocessor controlled photometer 96-well microplates MICROELISA  MICROTITER 5 different filters 405, 450, 490, 550, 595 (nm) Single and dual wavelengths eliminates some error.

ember
Download Presentation

Dynatech MR5000

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Dynatech MR5000 Using a Microplate Reader for a variety of tasks

  2. Features of the Dynatech MR5000 • Microprocessor controlled photometer • 96-well microplates • MICROELISA • MICROTITER • 5 different filters • 405, 450, 490, 550, 595 (nm) • Single and dual wavelengths • eliminates some error

  3. Applications • Kinetics • Agglutination • Tissue culture growth • Blood grouping • Complement fixation • MIC and bacterial ID assays • Advanced applications

  4. What I do with the MR5000 • Bradford Assay • total protein concentration of a sample • quick and easy • ELISA - more involved than Bradford • Enzyme-Linked Immunosorbent Assay • Buffers • 1 and 2 antibodies • Color Detection

  5. Bradford Assay • Homogenization & Centrifugation • Preparation of Samples & Standards • Plate Loading and Bradford Reagant • Coomassie Brilliant Blue G-250 & H3PO4

  6. What the Bradford tells us • Total protein concentration for each sample Why is this important? • To run ELISA need same amount of protein in each sample – for my work 2000 ng • For ELISA use a particular volume for each sample (determined by Bradford readings) FEB BRADFORDS completed.xls

  7. Monoclonal Antibody ELISA • Bradford data • Standards • Reagants • Binding Buffer (10mM Na2CO3/NaHCO3) • Blocking Solution – PBST (PBS + Tween 20) • PBST-BSA • Antibodies – • Primary • Secondary • Color Detection Solution • Diethanolamine R-nitrophenylphosphate tablets DAS-ELISA

  8. HSP 70

  9. Summary • So, we have: • Determined the concentration of protein in a sample (Bradford 595 nm) • Quantified the concentration of HSP 70 in equal volumes of protein (ELISA 405 nm) • Had a lot of fun!!

More Related