Navigation Links
Benchmark Plus Microplate Reader Scan Well Feature

Theresa Redila-Flores and Li Tian, PhD, Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules, CA 94547 USA


The Benchmark Plus microplate reader is a new monochromator- based absorbance reader from Bio-Rad that offers superior convenience, accuracy, and flexibility. The Benchmark Plus is controlled by Microplate Manager software. A new feature has been designed in Microplate Manager software (version 5.1 and higher) that utilizes the monochromator design of Benchmark Plus and allows the user to prescan a well in order to determine the optimal wavelength prior to running a full plate of samples. In this note, a prescan procedure is described to show how to implement such a function into real-life applications.

In the following procedure, standard protein (bovine IgG) is added to cell culture medium without serum and the Scan Well function in Benchmark Plus is used to determine the best wavelength for protein assay with cell culture medium. Bio-Rads protein assay kit (catalog # 500-0001) is used to check for measurement inconsistencies in culture medium.


Procedure
1. Serially dilute (1:2) standard bovine IgG protein ranging from 62.5 g/ml to 1 mg/ml in F-12 Hams medium without serum.

2. Aliquot 10 l (in triplicate) of each diluted standard protein into a 96-well clear bottom black microplate; i.e., A1, B1, and C1 for 1 mg/ml protein sample. Aliquot 10 l (in triplicate) of F-12 Hams medium into 3 wells (A7, B7, C7) to be used as blanks.

3. Add 200 l of diluted dye reagent (from Bio-Rad protein assay kit) to each well. Mix thoroughly using a microplate mixer for about 5 sec. Make sure that no bubbles are present. Bubbles may cause inconsistencies in absorbance reading.

4. Incubate 5 min at room temperature.

5. Place microplate into Benchmark Plus microplate reader.

6. Open Microplate Manager. Under File, select Endpoint protocol.

7. Select 96-well format. Under Advanced Options, open Scan Well.

8. Scan well A1 at 340800 nm using a 5 nm step size. Save data.

9. Scan remaining wells (A2A7) and save data.

10. For bovine IgG at optimal wavelength, select Endpoint protocol, 96-well format, and single wavelength reading at 590 nm.

11. Highlight the appropriate wells to be read; i.e., A1 to C7 for the concentration range from 62.5 g/ml to 1 mg/ml and the blank. 12. Read absorbance. Save and analyze data.


Results
The most concentrated protein sample shows a higher peak at 590 nm than the least concentrated protein samples (Figure 1). The least concentrated samples and the blank showed higher peaks at 470 nm, whereas the more concentrated protein samples showed similar peak heights at around 470 and 590 nm. From the figure, one can conclude that whereas a peak at 470 nm is apparently caused by the cell culture medium used here, the optimal wavelength for bovine IgG is at 590 nm.

Further reading of bovine IgG concentration at 590 nm on Benchmark Plus yields a linear graph with a correlation coefficient of 0.9993 (Figure 2).


Conclusion
The Benchmark Plus Scan Well feature is used to ascertain the best wavelength for reading a desired assay. It allows scanning of a well from 340800 nm at various wavelength increments. A graph and data of the scan area are provided and can be exported into Microsoft Excel software for further analysis. The Scan Well feature minimizes the problem of finding out at what wavelength samples should be read, thus saving time and effort. At the optimal wavelength for the bovine IgG tested, the correlation coefficient is ≥ 0.99 and linear over the dilution range of 62.5 g/ml to 1 mg/ml. The dilution in F-12 Hams medium without serum did not affect the measurement of the protein. The results show that the Benchmark Plus microplate reader is an excellent instrument to use for absorbance studies, as well as an instrument to save time in determining the optimal wavelength to read a sample.


Bibliography
Bradford MM, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem 72, 248254 (1976)

Lowry OH et al., Protein measurement with the Folin phenol reagent. J Biol Chem 193, 265275 (1951)


back to top
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Cell Proliferation Assay Using the Benchmark Plus Microplate Spectrophotometer
2. Benchmark Plus Microplate Reader: Quantitation of Protein Concentration Using Two Different Colorimetric Assay Kits, Rev A
3. Cerenkov Counting Performance on the TopCountTM Microplate Scintillation and Luminescence Counter
4. Counting Non-Aqueous Samples in the TopCount Microplate Scintillation and Luminescence Counter
5. Fluorometric Protease Assays in the SpectraMax Gemini Microplate Spectrofluorometer: Example Using Caspase3 (MaxLine Application Note #35)
6. LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43)
7. Using the OligoGreen Oligonucleotide Quantitation Reagent in the Gemini XS, Gemini EM and SpectraMax M2 Microplate Readers (MaxLine Application Note #21)
8. PathCheck Applied to Measurement of Protein Solutions in the SpectraMax Plus Microplate Spectrophotometer (MaxLine Application Note #26)
9. Microdetermination of Phosphorus Using the SpectraMax Plus Microplate Spectrophotometer: Choice of Microplate, Cuvette, or Test Tube Assay Formats (MaxLine Application Note #24)
10. UV Absorbance Measurements in SpectraMax Microplate Spectrophotometers MaxLine Application Note #32)
11. Ca++-Induced G Protein Coupled Receptor (GPCR) Activation Monitored via Aequorin Luminescence in the LMax Microplate Luminometer (MaxLine Application Note #42)
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Benchmark Plus Microplate Reader Scan Well Feature

(Date:7/10/2014)... -- According to the International Atomic Energy Agency ... from terrorists acquiring sufficient quantities of plutonium or ... nuclear explosive device. The IAEA also notes that ... gram-level quantities, which can be challenging to detect ... new study appearing this week in the ...
(Date:7/10/2014)... 2014 Using microscopic polymer light resonators that ... at MIT,s Quantum Photonics Laboratory have developed new ... parts-per-billion range. Optical sensors are ideal for detecting ... ratio, compact, lightweight nature, and immunity to electromagnetic ... been developed before, the MIT team conceived an ...
(Date:7/10/2014)... of psychiatric disorders has been extremely challenging because there ... are insufficient to cause disease. Now investigators reporting in ... Cell Stem Cell describe a strategy that ... with other risk factors or environmental exposures to affect ... a genetic variant that may predispose individuals to schizophrenia. ...
(Date:7/10/2014)... batteries that power our laptops and electric vehicles could ... charge with the help of a sponge-like silicon material. ... the graphite traditionally used in one of the battery,s ... energy storage capacity of graphite. A paper describing the ... today in Nature Communications . , "Silicon has ...
Breaking Biology Technology:Sensitive detection method may help impede illicit nuclear trafficking 2Swell new sensors 2New strategy could uncover genes at the root of psychiatric illnesses 2Silicon sponge improves lithium-ion battery performance 2Silicon sponge improves lithium-ion battery performance 3
... adison, Wis. - Third Wave Technologies, Inc. ... a share, for the second quarter of 2006, but robust ... upward in a key market. , ,Third Wave, which develops ... that revenue rose by 15 percent for the quarter ended ...
... Madison, Wis. - A United States Air Force contract ... not only provide a Madison aerospace research and product development ... it to propel something much larger. , ,The U.S. ... Orbital Technologies Corp. (Orbitec) for the development of ...
... - Thanks to a classified contract from the ... company headquartered here, reported record revenues and earnings for ... the closing of its manufacturing facility in England. , ... of lower-than-expected earnings in the fourth quarter, which caused ...
Cached Biology Technology:Third Wave reports another loss, stronger sales 2Third Wave reports another loss, stronger sales 3Orbitec will use $25 million Air Force contract to make small launch vehicle 2Defense contract helps Plexus gain record profits 2
(Date:7/11/2014)... the Department of Energy,s Oak Ridge National Laboratory ... R&D Magazine in recognition of the year,s top ... tremendous value of our National Labs," said Secretary ... the National Labs continues to help our nation ... and technological innovations necessary to remain globally competitive." ...
(Date:7/11/2014)... The University of Chicago is creating a new ... at the University,s Institute for Molecular Engineering and ... million donation from the Millicent and Eugene Bell ... will reside within the Institute for Molecular Engineering. ... research project at the MBL,s Eugene Bell Center ...
(Date:7/11/2014)... The Chinese University of Hong Kong, BGI and other ... to salt tolerance, with implication for improving this important ... online in Nature Communications provides an effective ... , Soybean is an important crop for the world. ... less genetic diversities than their wild counterparts. Among the ...
Breaking Biology News(10 mins):ORNL wins eight R&D 100s 2ORNL wins eight R&D 100s 3ORNL wins eight R&D 100s 4New professorship in tissue engineering links molecular engineering, marine biology 2BGI reports a novel gene for salt tolerance found in wild soybean 2
... the University of Leeds are developing methods of helping ... chemicals such as vinegar. , The harmful chromium ... from former textiles factories, smelters, and tanneries have been ... problems with the kidneys, liver, lungs and skin. , ...
... but researchers in China have discovered that chicken manure can ... in the International Journal of Environment and Pollution ... 50% more crude oil than soil lacking the guano. ... University, China, point out that contamination of soil by crude ...
... comes to solving complex problems, Geoffrey von Maltzahn, MIT ... solutions. Finding inspiration in systems that evolution has produced, ... society,s biggest challenges: improving tumor detection and therapeutic delivery ... patients. Today, the 28-year-old Ph.D. candidate in ...
Cached Biology News:Cleansing toxic waste -- with vinegar 2Fowl soil additive breaks down crude oil 2MIT student develops new innovations to selectively kill cancer cells 2MIT student develops new innovations to selectively kill cancer cells 3MIT student develops new innovations to selectively kill cancer cells 4
Mouse monoclonal [3G131] to Desmoglein 1 ( Abpromise for all tested applications). entrezGeneID: 1828 SwissProtID: Q02413...
Mouse monoclonal [2D7/1] to E.coli LPS ( Abpromise for all tested applications). Antigen: Tissue / cell preparation: Escherichia coli J5 cells....
COMING SOON: Anti-Mouse C5/C5b, Clone BB5.1. Monoclonal Antibody...
KAI 1 (G-2)...
Biology Products: