Buffer Preparation Lab Report
Procedure:
Day 1: Buffer preparation
First, the buffer was prepared by using the formula as follows:
Figure 1: Calculation for prepare 0.5 M Tris buffer at pH 6.8
3.033 g of Tris was weighed and placed in 400 mL beaker. Then, 25 mL of distilled water was added into the beaker that contained Tris. The mixture was dissolved using the stirring rod, and then the magnetic stirring bar was placed in the beaker for further dissolve when measuring the pH. The pH meter was used to measure the solution, and the data was documented at pH 11.40. This was the starting point. Next, 6M HCl was slowly added into the buffer to make it to pH 6. Then, the buffer was transferred to a 100mL graduated cylinder, and 25mL of distill water was added to the buffer. Next, the buffer was transferred to 125 mL glass jar and was labeled 0.5M Tris buffer, pH 6.8. The top of the flask was sealed with Parafirm and was placed in the refrigerator for next day lab. Other group was prepared other buffers that needed for this experiment. They are 0.5M Tris pH 6.8, 1x-running buffer, and transfer buffer.
Day 2: Buffer preparation and run the gel and the samples
First, the buffer was prepared by using the formula as follows: …show more content…
The primary antibody recognizes a specific amino-acid sequence of a particular protein. In this case, the antibody was anti-cow albumin. Then, it was incubated at 37°C for 30 minutes. After incubated, it was rinsed with TBS +NP 40 1x for 4 times. Each time was 5 minutes. This step was to wash the unbound antibodies. Then, it was rinsed two times with TBS 1x for 5 minutes to remove the soap. The blot was then developed using the color development. 7mL of chlornaptol and 1μL H2O2 was measured and added into the nitrocellulose and it was allowed to sit for 10-15 minutes for the color to develop. Then, the light purple color was presented. The picture was taken and recoded in
Day 1: Buffer preparation
First, the buffer was prepared by using the formula as follows:
Figure 1: Calculation for prepare 0.5 M Tris buffer at pH 6.8
3.033 g of Tris was weighed and placed in 400 mL beaker. Then, 25 mL of distilled water was added into the beaker that contained Tris. The mixture was dissolved using the stirring rod, and then the magnetic stirring bar was placed in the beaker for further dissolve when measuring the pH. The pH meter was used to measure the solution, and the data was documented at pH 11.40. This was the starting point. Next, 6M HCl was slowly added into the buffer to make it to pH 6. Then, the buffer was transferred to a 100mL graduated cylinder, and 25mL of distill water was added to the buffer. Next, the buffer was transferred to 125 mL glass jar and was labeled 0.5M Tris buffer, pH 6.8. The top of the flask was sealed with Parafirm and was placed in the refrigerator for next day lab. Other group was prepared other buffers that needed for this experiment. They are 0.5M Tris pH 6.8, 1x-running buffer, and transfer buffer.
Day 2: Buffer preparation and run the gel and the samples
First, the buffer was prepared by using the formula as follows: …show more content…
The primary antibody recognizes a specific amino-acid sequence of a particular protein. In this case, the antibody was anti-cow albumin. Then, it was incubated at 37°C for 30 minutes. After incubated, it was rinsed with TBS +NP 40 1x for 4 times. Each time was 5 minutes. This step was to wash the unbound antibodies. Then, it was rinsed two times with TBS 1x for 5 minutes to remove the soap. The blot was then developed using the color development. 7mL of chlornaptol and 1μL H2O2 was measured and added into the nitrocellulose and it was allowed to sit for 10-15 minutes for the color to develop. Then, the light purple color was presented. The picture was taken and recoded in