Determining the Protein Content of Nuclear Pellets from Kidneys of Rats - Lab Report Example

The concentration of protein was determined by the use of a calibration curve. The experiment used Peterson’s modification of the micro-Lowry technique that precipitates the protein by the use of deoxycholate (DOC) and trichloroacetic acid (TCA) to avoid interferences from other chemicals. The precipitation was carried out in Laboratory 6.     

Methods

The following method was used in the preparation of Kidney Nuclear Pellets from the Cortex and medulla for protein determination.

Detailed procedure

Part 1

1. The recording of the samples to be prepared on the attached sheet was done.
2. The sample tubes and the pellet were weighed and their weights recorded.
3. 0 ml of 0.1 M NaOH was added to each tube and reweighing done.
4. The tubes were shaken until the pellets dissolved.

Part 2

1. The tubes were centrifuged at 10,000 rpm for 10 minutes in order to remove the turbidity from the solution, making sure that the tubes were not disturbed or shaken prior to pipetting.
2. In every sample, there was the labeling of 2 clean microcentrifuge tubes with the same sample ID. The tubes then became assay tubes.
3. 150 ul of the original sample was added to the duplicate assay tubes.
4. 450 ul of distilled water was added to the assay tubes after which they were shaken.
5. 100 ul of DOC solution was added to all the tubes and mixed and then allowed to stand for 10 minutes.
6. 100 ul of TCA solution was added to all the tubes and mixed.
7. All the tubes were centrifuged for 10 minutes at 10, 000 x g.
8. The supernatant was removed.

Result

The result for CY41CNAfB was obtained. The pellet was preserved for use in laboratory 7.

Conclusion

There was a successful dilution of the sample from ratio one to four. The results of the study were plotted on a graph for protein determination. It was concluded that the protein concentration in the kidney cortex and medulla of rats was lower than that of other samples.