Question

Western Blot of LDH:

1. Suppose, upon developing your blot, you see more bands that you expect. What are two possible reasons for this?

2. Suppose, upon developing your blot, you see no bands at all. What are two possible reasons for this?

Answer 1

Answer 1:

for more bands as compare to expectations is due to:

• Impurities of proteins (mainly) or other content in sample used for blotting.
• Sometimes impurity in antibody or other expiry content can affect the analysis and banding patterns.
• Handling error can also generate unwanted bands.
• Unwanted or unorganized binding of antibodies.

Answer 2:

for less bands as compare to expectations is due to

• Same conditions: Improper sample loading in gel and Impurities of proteins (mainly) or other content in sample used for blotting.
• Improper centrifugation of protein and handling error.
• Incomplete binding of antibodies and protein.
• Biological reaction in cell leads to change in product