Anti-flag Agarose
Anti-flag Agarose - Related Videos
13:46
Immunoprecipitation-Based Techniques: Purification of Endogenous Proteins Using Agarose Beads
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2023
Source: Susannah C. Shissler1, Tonya J. Webb1 1 Department of Microbiology and Immunology, University of Maryland, Baltimore, MD 21201 Immunoprecipitation (IP, also known as a 'pull-down' assay) is a widely used technique that has applications in a variety of fields. First conceived in 1984, it was refined in 1988 (1, 2). The fundamental goal of IP is purification and isolation of a specific protein using an antibody against that protein. The word "immuno" refers to the use of an antibody while...
09:24
Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins
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Cited by 32 •
2016
Mucins are high-molecular-weight glycoconjugates, with size ranging from 0.2 to 200 megadalton (MDa). As a result of their size, mucins do not penetrate conventional polyacrylamide gels and require larger pores for separation. We provide a detailed protocol for mucin agarose gel electrophoresis to assess relative quantification and study polymer assembly.
04:26
Agarose-Based Preparation of Human Precision-Cut Lung Slices
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2025
In this video, we describe a procedure to inflate isolated human lungs with agarose and prepare human precision-cut lung slices. The prepared ex vivo lung slices can be used for further immunologic experiments.
07:53
Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
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Cited by 8 •
2019
In this procedure, a DsRed-based epitope ligand is immobilized to produce a highly selective affinity resin for the capture of monoclonal antibodies from crude plant extracts or cell culture supernatants, as an alternative to Protein A.
02:35
DNA Agarose Gel Electrophoresis
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2021
Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol. Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA. In cloning experiments, both the insert and vector DNA...
FAQs
Selectivity comes from the interaction between immobilized anti-FLAG antibodies and the engineered FLAG epitope. Because the antibodies are attached to the agarose support, the target protein can be retained while many other molecules in a cell lysate remain unbound. This molecular recognition is the basis for enriching a tagged protein from a complex biological sample.
Washing separates molecules that did not bind specifically from the antibody-resin complex. During this step, unbound proteins and other sample components are removed, whereas FLAG-tagged material remains associated with the immobilized antibodies. The effectiveness of this separation determines how much background is reduced before recovery of the enriched protein fraction.
Recovery can be achieved by introducing excess FLAG peptide or by changing the binding conditions. Excess peptide can help release the captured protein, whereas altered conditions can disrupt the binding environment. These alternatives give researchers different ways to separate the target from the resin after other sample components have been washed away.
A basic workflow starts with a cell lysate or another complex sample, followed by contact with the resin so FLAG-tagged proteins can bind. The resin is then washed to remove unbound material, and the retained proteins are recovered through controlled elution. The resulting fraction is enriched for the tagged protein and suited to downstream analysis.
In molecular and cellular biology, this resin is useful when a study requires an enriched FLAG-tagged protein rather than the full mixture present in a lysate. The recovered material can support protein purification and detection, allowing investigators to examine the target after isolation. It also provides a practical starting sample for biochemical assays and protein characterization.
Interaction studies can use the enriched fraction to examine proteins associated with the FLAG-tagged target, while functional analysis can focus on the isolated material in a biochemical assay. Detection provides an additional way to assess whether the tagged protein was recovered. Together, these uses connect affinity-based enrichment with questions about molecular associations, protein function, and biological activity.