E11: protein gel electrophoresis

1. aspartic acid

2. glutamic acid

acidic amino acids

1. lysine

2. Arginine

3. Histidine

Basic amino acids

true

Any amino acid can be charged under appropriate condition. True or False?

pI

the point at which the protein does not migrate in an electric field (isoelectric point)

pI > pH

positively charged

moves toward cathode (-)

pI < pH

negatively charged

moves to the anode (+)

sickle cell

an abnormal red blood cell that has a crescent shape and an abnormal form of hemoglobin

has a distinct two fewer negative charges. This means that it would move less distance to the anode compared to normal.

the difference of the pI and the pH. the larger the difference the more positive/negative

What determines the direction of movement of the proteins?

no

Does the molecular weight of a protein affect its migration?

no

Does the separation require proteins to be denatured?

no

Does the protein change its shape while it is moving in the gel?

outside

Primers need to be designed

false, it only recognize human cell

primer design PCR can anneal to DNA of both human epithelial cells and bacteria collected in mouthwash

true

The isoelectric point (pI) of an amino acid is defined as the pH at which the amino acid does not migrate in an electric field or the pH at which an amino acid bears no net charge.

bovine serum albumin (pI = 4.8)

Which of the following four proteins in Part A of today's experiment is the most acidic protein?

cation, cathode

If amino acid Lysine with a pI of 9.7 is dissolved in a gel running buffer of pH 6; at this pH, Lysine is a___________ and it will migrate toward____________ during the native agarose gel electrophoresis

net charge difference at a particular pH

What is the basis of separation of the HbA and HbS in Part B of today's experiment?

- Separation of DNA molecules using gel electrophoresis is on the basis of sizes

-Separation of Protein using "Native" or "non-denaturing" gel electrophoresis is on the basis of the net charges

Which two of the following statements about the separation of molecules are true?

- 0.72 g of agarose

- 60 ml of electrophoresis buffer with pH at 9.2

If your group is assigned to prepare 60 ml of 1.2% agarose gel solution for part B, what materials are required to prepare the gel solution. Select the amount of agarose and the amount of the correct buffer needed