Cara Pemeriksaan Protein Total

Summary

This video script details a laboratory procedure for testing total protein levels using specific reagents. The process includes preparing the monoreagent by mixing two reagents in a precise ratio, followed by sample preparation in cuvettes for reaction. After incubation, the protein sample's absorbance is measured using a spectrophotometer. The script outlines the step-by-step process of adding reagents, mixing, and reading absorbance values to calculate the total protein concentration. It emphasizes precision in measurements and careful handling during the procedure to ensure accurate results.

Takeaways

• 😀 Preparation of the monoreagen involves mixing 4 mL of Reagen 1 with 1 mL of Reagen 2 in a test tube.
• 😀 The monoreagen must be mixed 3-5 times to ensure proper blending and avoid bubble formation.
• 😀 Three cuvettes are prepared: one for the sample, one for the standard, and one for the blank control.
• 😀 20 mL of distilled water is added to the blank cuvette for calibration.
• 😀 A 20 µL sample of the protein is added to the cuvette designated for the sample.
• 😀 1000 µL of the monoreagen is added to the sample cuvette and gently mixed 3-5 times.
• 😀 The same process is followed for the standard protein sample in a separate cuvette.
• 😀 The samples are incubated for 5 minutes to allow the reaction to occur, resulting in a visible layer.
• 😀 Absorbance is measured using a spectrophotometer set at 546 nm wavelength.
• 😀 The blank cuvette is set as the baseline before measuring the absorbance of both the standard and the sample.
• 😀 The absorbance readings are used to calculate the total protein concentration based on a standard curve formula.

Q & A

What is the first step in preparing for the total protein test?

The first step is to prepare the mono reagent by mixing 4 parts of reagent 1 with 1 part of reagent 2. This mixture is then placed in a reaction tube.

How is the mono reagent prepared?

The mono reagent is prepared by pipetting 4 milliliters of reagent 1 into a reaction tube, then adding 1 milliliter of reagent 2. The mixture is then mixed thoroughly 3-5 times.

What is the purpose of preparing the mono reagent?

The mono reagent is essential for conducting the total protein test. It helps in reacting with the sample to measure the total protein concentration.

What equipment is used for the protein test?

A spectrophotometer is used to measure the absorbance during the protein test, and cuvettes are used to hold the samples.

How is the sample prepared for measurement?

20 microliters of the sample is placed in a cuvette, followed by the addition of 1000 microliters of the prepared mono reagent. The sample is then mixed gently to avoid bubbles.

What is the importance of mixing the sample properly?

Proper mixing is essential to ensure the reagent reacts uniformly with the sample, allowing for accurate absorbance readings. Care must be taken to avoid bubbles.

How long should the sample be incubated?

The sample should be incubated for 5 minutes before proceeding with the absorbance measurement.

At what wavelength is the absorbance measured?

The absorbance is measured using a spectrophotometer set at a wavelength of 546 nanometers.

What is the purpose of using a blank in this experiment?

A blank is used to calibrate the spectrophotometer and ensure accurate measurements by setting a baseline absorbance.

How is the protein concentration calculated?

The total protein concentration is calculated using the absorbance values obtained for both the standard and the sample, applying a specific calculation formula.