Western Blotting: A Key Technique for Protein Detection and Analysis
35. What are the techniques use to transferred protein onto nitrocellulose paper?
a. Far western
b. Southern blotting
c. Northern blotting
d. Western blotting
Understanding Western Blotting (Western blotting Protein transfer)
Western blotting is a widely used laboratory technique in molecular biology and biochemistry to detect and analyze specific proteins in a complex mixture. It involves separating proteins, transferring them onto a membrane, and detecting them using antibodies. The most commonly used membrane for this process is nitrocellulose or PVDF (polyvinylidene difluoride).
Why Transfer Proteins onto Nitrocellulose Paper?
After separating proteins via SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis), they need to be transferred onto nitrocellulose paper because:
- Proteins remain immobilized on the membrane.
- Easy antibody binding for detection.
- Enhanced protein stability for prolonged analysis.
Western Blotting: The Correct Answer
Steps in Western Blotting:
- Protein Separation – Proteins are separated using SDS-PAGE based on molecular weight.
- Transfer onto Nitrocellulose – Proteins are transferred from the gel to nitrocellulose paper using an electric field (electroblotting).
- Blocking – The membrane is blocked using non-fat milk or BSA to prevent non-specific binding.
- Primary Antibody Binding – A specific antibody binds to the target protein.
- Secondary Antibody Binding – A labeled secondary antibody binds to the primary antibody for detection.
- Detection – The protein of interest is visualized using chemiluminescence, fluorescence, or colorimetric methods.
Why Other Options Are Incorrect
-
Far Western Blotting (Option A):
- A specialized form of Western blotting that detects protein-protein interactions.
- Not a standard method for transferring proteins.
-
Southern Blotting (Option B):
- Detects DNA fragments, not proteins.
-
Northern Blotting (Option C):
- Detects RNA sequences, not proteins.
Thus, the correct answer is:
✅ (D) Western Blotting
Conclusion
Western blotting is the gold standard technique for transferring and detecting proteins on nitrocellulose membranes. It plays a crucial role in biomedical research, disease diagnosis, and protein characterization, making it a fundamental tool in molecular biology.
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70 Comments
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Khushi Vaishnav
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August 20, 2025(D) Western Blotting
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August 20, 2025Ans d
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Diksha Chhipa
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August 20, 2025Answer d Western blotting are the technique used to transferred protein into nitrocellulose membrane
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August 20, 2025The correct answer is option 4th
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August 21, 2025Option D is correct
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Mitali saini
August 23, 2025Western Blotting: The Correct Answer Proteins remain immobilized on the membrane.
Easy antibody binding for detection.
Enhanced protein stability for prolonged analysis.
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August 23, 2025Option d
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August 30, 2025Correct Answer is option (d )
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