Protein Determination Laboratory Report

Categories: Biology

Report, Pages 3 (688 words)

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Introduction

Proteins play a vital role in the growth and maintenance of the human body, serving as essential components comprised of lipids and carbohydrates. This laboratory experiment employs the Bradford assay to analyze the levels of Albumin in patient urine samples, a crucial diagnostic tool for assessing protein concentration and identifying abnormalities in patients.

Albumin, constituting approximately 70% of plasma protein, is primarily synthesized in the liver and serves to maintain a concentration gradient between blood and extracellular tissues. Normal albumin levels in a healthy individual typically range from 20ug/ml to 30ug/ml.

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Deviations from this range, such as albumin levels exceeding 30ug/ml, may indicate conditions like microalbuminuria, which elevates the risk of cardiovascular diseases, including strokes and hypertension.

The laboratory analysis of abnormal albumin concentrations in patient urine samples is essential for diagnosing conditions related to kidney damage, eclampsia, and diabetes. Proteinuria, the presence of protein in urine, is often a marker of kidney damage, with individuals exhibiting high levels of albumin in urine being at an increased risk of cardiovascular diseases due to elevated blood pressure.

Research has shown that proteinuria can affect both men and women, whether or not they have diabetes, emphasizing its clinical significance.

Methodology

For this laboratory experiment, the Bradford Assay was employed using Coomassie blue dye G-250. Eight standard tubes were used, with protein concentrations ranging from 0ul (standard 0) to 1000ul (standard 7).

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Bovine serum Albumin (BSA) at a concentration of 1mg/ml (1µg/µl) was added to labeled microtubes, and 100 µl of each standard was duplicated and mixed with 5 ml of diluted Bradford reagent.

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The mixture was left at room temperature for approximately 10 minutes, followed by recording the absorbance at 595 nm using a spectrometer. The absorbance values were then recorded in an Excel spreadsheet to create a calibration graph. The calibration graph was used to analyze the protein concentration of patient samples.

Each patient sample, containing 100µl of the standard and 5ml of Bradford reagent, was also read at 595nm to record absorbance values. The average absorbance values were used to quantify the albumin protein concentration in each patient sample.

Results

The results of the laboratory experiment are summarized in the following tables:

Table 1: Calibration Table of Range of Albumin Concentration

Albumin Concentration (ug/ml)	Average Absorbance @595nm
0	3.25E-2
20	5.95E-2
50	0.1035
75	0.1425
100	0.3235
250	0.8655
500	1.0

Table 2: Quantification of Albumin Concentrations (ug/ml) in Patient Samples and Controls with Standard Deviation

Concentration	Patient 1	Patient 2	Patient 3	Patient 4	Patient 5	Normal Control	High Control
Patient Urine Samples	116.58	96.14	61.23	178.10	44.27	20	250

Discussion

In a healthy urine sample, normal albumin concentrations should be exceptionally low, ranging from 30 ug/ml to 80 ug/ml.

The analysis of patient samples revealed the following:

Patient 1 - 35-year-old Female: Abnormally high protein concentration of approximately 120 ug/ml, suggesting the possibility of pre-eclampsia, a condition characterized by elevated blood pressure during pregnancy. Further blood tests are recommended to assess kidney and liver function.

Patient 2 - 50-year-old Male: Albumin concentration of approximately 99 ug/ml, indicating microalbuminuria. This patient is overweight and exhibits hematuria, possibly due to the presence of small kidney stones.

Patient 3 - 40-year-old Female: Albumin concentration of approximately 52ug/ml, within the normal range (50-80 ug/ml).

Patient 4 - 23-year-old Female: Significantly elevated albumin concentration of approximately 170 ug/ml, suggestive of albuminuria. The patient presents symptoms of Urinary Tract Infection.

Patient 5 - 70-year-old Male: Albumin concentration of around 48 ug/ml, similar to the normal control sample. The patient exhibits flu-like symptoms, which may be attributed to a weakened immune system due to age.

Calibration Table of Range of Albumin Concentration

Albumin Concentration (ug/ml)	Average Absorbance @595nm
0	3.25E-2
20	5.95E-2
50	0.1035
75	0.1425
100	0.3235
250	0.8655
500	1.0

Quantification of Albumin Concentrations (ug/ml) in Patient Samples and Controls with Standard Deviation

Concentration	Patient 1	Patient 2	Patient 3	Patient 4	Patient 5	Normal Control	High Control
Patient Urine Samples	116.58	96.14	61.23	178.10	44.27	20	250

Conclusion

The laboratory experiment successfully quantified albumin concentrations in patient urine samples, allowing for the identification of abnormal levels indicative of various health conditions. The results indicated microalbuminuria in several patients, suggesting the need for further medical evaluation.

Early detection of abnormal albumin concentrations is crucial for preventing complications such as hypertension, diabetes, and heart disease. The diagnostic value of protein determination in urine samples is evident, providing valuable insights into patients' health.