importance of the standard curve and spectrophotometry

importance of spectrophotometry and the standard curve

We hypothesized that the different wavelengths, absorbance, and concentration of a known protein (one percent albumin) will help us find an unknown concentration by using the standard curve of the known serial diluted concentration.

by: my-toast-is

Sample Preparation

Peak Wavelength

The importance of the Standard curve

how can we improve our experiment and how can it benefit our society?

tubes 1-5

tubes U AND B

(Elisa Micro Plate Reader)

• ELISA it's micro plate reader that emits light using one wave length and measures the amount of light absorbed which is reflected by an object such as a protein or a known substances.
• The spectrophotometer measures the conversion of a clear substance to a colored product by the coupled enzyme.
• The spectrophotometer is important in the ELISA experiment because you can see the readings from one particular protein or substance versus a bunch. Therefor allowing you to concentrate on just one known substance.

absorbance

Introduction

Spectrophotometry

[spek-troh-foh-tom-i-ter]

Spectroscopy

Spectrophotometry

[spek-troh-foh-tom-i-tree]

A method to measure how much a chemical substance absorbs light by measuring the intensity of the light as a beam of light passes through the sample solution.

SPECTROSCOPY

the branch of science concerned with the investigation and measurement of spectra produced when matter interacts with or emits electromagnetic radiation.

Standard Curve

It is used to determine the concentration of a known substance.

Serial Dilution

Is a laboratory technique in which a substance is decreased in a concentration of a series of equal measurements. A solution sample may be distributed in a series of tubes so that each has one half of the amount of the previous tube in the series.

Absorbance

a measure of the capacity of a substance to absorb light of a specified wavelength. It is equal to the logarithm of the reciprocal of the transmittance.

wavelength

The distance between peaks (high points).

Standard curve

Serial dilution

Outline

Absorbance

wavelength

1. Introduction IV. Results

II. Hypothesis V. Conclusion

III. Methods VII. Broader Impact

Questions?

x= (y-b)/m

In conclusion, we found that the spectrophotometer is a very useful tool in finding out exact measurements for a single substance. Therefore, allowing us to know the absorbance of the serial diluted concentration that we tested.

questions

Background Information

• The spectrophotometer is useful because the absorbance and transmittance of light energy and is the basis for a widely used procedure for determining the concentration of substances in a solution.

• This lab includes the principles of the Scientific Method, a series of steps commonly used to make observations, formulate hypothesis, carry out experiments and interpret the data generated from these experiments.

REFERENCES

• Enzyme-Linked Immunosorbent Assay (ELISA) - Boundless Open Textbook. (n.d.). Retrieved April 21, 2016, from https://www.boundless.com/microbiology/textbooks/boundless-microbiology-textbook/immunology-applications-12/diagnostic-immunology-148/enzyme-linked-immunosorbent-assay-elisa-748-10812/

• Microplate reader vs spectrophotometry - the advantages and disadvantages (Jul/19/2007 ). (n.d.). Retrieved April 21, 2016, from http://www.protocol-online.org/biology-forums/posts/29037.html

• Smith, B. (n.d.). What Is an ELISA Reader? Retrieved April 21, 2016, from http://www.ehow.com/facts_7573969_elisa-reader.html

References

• Enzyme-Linked Immunosorbent Assay (ELISA) - Boundless Open Textbook. (n.d.). Retrieved April 21, 2016, from https://www.boundless.com/microbiology/textbooks/boundless-microbiology-textbook/immunology-applications-12/diagnostic-immunology-148/enzyme-linked-immunosorbent-assay-elisa-748-10812/

• Microplate reader vs spectrophotometry - the advantages and disadvantages (Jul/19/2007 ). (n.d.). Retrieved April 21, 2016, from http://www.protocol-online.org/biology-forums/posts/29037.html

• Smith, B. (n.d.). What Is an ELISA Reader? Retrieved April 21, 2016, from http://www.ehow.com/facts_7573969_elisa-reader.html

• T. (2014). What is a Standard Curve? Retrieved April 22, 2016, from youtube/Whatisastanardcurve