The Protein Man's Blog | A Discussion of Protein Research

How to Prepare Samples for Western Blot Analysis

Posted by The Protein Man on Sep 8, 2020 9:00:00 AM

What is Western Blot Analysis?

Western blotting or immunoblotting is an indispensable technique, almost every published paper in area of molecular cell biology uses western blotting for detecting specific proteins in a sample of tissue homogenate or cell lysates. Western blotting combines resolving power of polyacrylamide gel electrophoresis (PAGE) or SDS-PAGE and specificity of antibodies to detect target proteins.  Proteins are resolved on the basis of their molecular weight in SDS-PAGE and transferred from the polyacrylamide gel onto the membranes (Nitrocellulose or PVDF), which creates an exact replica of the protein separation pattern on the membrane. After transferring the proteins to the membrane, the membrane needs ‘blocking’ to ‘block’ non-specific binding sites on the surface of the membrane. Blocking is usually performed with Bovine Serum Albumin, Skimmed milk or purified milk Casein.

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The era of messenger RNA vaccines

Posted by The Protein Man on Aug 28, 2020 10:00:00 AM

“Trumping the vaccine game”

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Guess what??? Immobilized Papain Generates Fab Fragments of Cluster A Monoclonal Antibodies for Validation of Inner Domain of HIV-1 gp120?

Posted by The Protein Man on Aug 19, 2020 3:15:00 PM

Introduction

The lab of Dr. William D. Tolbert and Dr. Neelakshi Gohain use Papain-linked agarose resin to create combinations of purified Fabs of Cluster A monoclonal antibodies (A32 mAbs and JR4 mAbs) to validate the design and crystal structure of a stabilized inner domain (ID) of HIV-1 gp120. This ID displays a major antibody-dependent cellular cytotoxicity (ADCC) target of the A32 region. Current research supports a role for ADCC in preventing HIV-1 infection and in vaccine-induced protection that correlates with slower HIV progression or decreased virus replication.

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Topics: Molecular Biology, Protein Estimation, Protein Concentration, Protein Labeling, Buffers & Chemicals

What is the Link between Protein Folding and Prion Disease?

Posted by The Protein Man on Aug 12, 2020 5:00:00 PM



Prion diseases (also referred to as transmissible spongiform encephalopathies, or TSEs) are a group of progressive, untreatable, and fatal neurodegenerative conditions affecting both humans and animals. Generally, these conditions are characterized by long incubation periods (usually between 5 to 20 years), disruption in the normal tissue structure (resulting in holes and vacuole formation in the neurons), and the absence of an inflammatory response.

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Topics: Protein Purification, Molecular Biology, Western Blotting, Protein Electrophoresis, Cytotoxicity Assays, Sample Clean Up, Protein Concentration, Protein Fractionation, Protein Labeling, Protein Extraction, Buffers & Chemicals

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