Weetern Blot Flashcards
(24 cards)
What is the primary purpose of a Western blot?
To detect specific proteins in a sample.
True or False: A clear band on a Western blot indicates the presence of protein regardless of the loading control.
False
Fill in the blank: The _____ control is essential for validating the amount of protein loaded in each lane of a Western blot.
loading
Which of the following factors can affect the interpretation of Western blot results? A) Antibody specificity B) Sample preparation C) Transfer efficiency D) All of the above
D) All of the above
What is one critical analysis step to consider when evaluating the bands on a Western blot?
Comparing the band intensity to a standard or control.
True or False: Western blotting can quantify the amount of protein present in a sample.
True.
What does a band on a Western blot represent?
The presence and size of a specific protein.
Fill in the blank: The blocking step in a Western blot is performed to prevent __________.
Non-specific binding.
Multiple Choice: Which of the following factors can affect the results of a Western blot? A) Antibody specificity B) Sample loading amount C) Transfer efficiency D) All of the above
D) All of the above.
What does it mean if a Western blot shows multiple bands for a single protein?
It may indicate post-translational modifications or protein isoforms.
Fill in the blank: Western blot results should always be compared to a __________.
Molecular weight marker.
What is the significance of using a loading control in Western blots?
To normalize the protein levels across different samples.
What can cause high background noise in a Western blot?
Excessive primary antibody concentration or insufficient washing.
Multiple Choice: Which of the following can lead to false positives in Western blots? A) Cross-reactivity B) Improper washing C) Overexposure D) All of the above
D) All of the above.
True or False: The results of a Western blot can be influenced by the type of sample used.
True.
Fill in the blank: To improve reproducibility, it is essential to keep __________ between experiments.
All conditions consistent.
What can cause smearing in a western blot?
Overloading the gel, incomplete protein denaturation, or poor transfer efficiency.
Fill in the blank: Smearing can obscure the _____ of the target protein in a western blot.
bands.
What is one method to reduce smearing in western blots?
Optimizing sample concentration.
What is a common indicator of smearing in a western blot?
Broad bands instead of sharp, discrete bands.
How can sample preparation affect smearing in a western blot?
Improper lysis or denaturation can lead to protein aggregation.
What does the term ‘background noise’ refer to in the context of western blotting?
Unwanted signals that can obscure the target protein bands.
What is the effect of using too much sample volume in a western blot?
It can lead to band smearing and poor resolution.
Multiple Choice: What type of gel is commonly used in western blotting?
SDS-PAGE gel.
