FLAG-HRP Antibody
Description
Antibody Types and Specificity
FLAG-HRP antibodies are available as monoclonal or polyclonal, with variations in host species and isotypes:
Monoclonal antibodies (e.g., M2 clone) are often preferred for their consistent performance, while polyclonal antibodies offer broader epitope recognition .
Applications and Protocols
FLAG-HRP antibodies are versatile tools in molecular research:
Western Blotting
ELISA
Immunoprecipitation and Immunocytochemistry
Research Insights and Optimization
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Sensitivity: Monoclonal antibodies like M2 exhibit high specificity, detecting as little as 10 µg of lysate .
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Compatibility: Hydrophilic FLAG tags minimize interference with protein folding or function .
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Troubleshooting: Overblocking with milk/gelatin or inadequate substrate exposure can reduce signal; optimize dilutions and exposure times .
Product Specs
Product Science Overview
The FLAG peptide is a short, hydrophilic peptide consisting of eight amino acids: Aspartic acid (D), Tyrosine (Y), Lysine (K), Aspartic acid (D), Aspartic acid (D), Aspartic acid (D), Aspartic acid (D), and Lysine (K), represented by the sequence DYKDDDDK . This peptide is widely used as a fusion tag for the purification and detection of recombinant proteins.
The FLAG peptide is specifically designed for immunoaffinity chromatography, allowing for the elution of fusion proteins under non-denaturing conditions . It is commonly used in various biochemical and molecular biology techniques, including:
- Western Blotting: For detecting FLAG-tagged proteins.
- Immunocytochemistry: For visualizing the localization of FLAG-tagged proteins within cells.
- Immunoprecipitation: For isolating FLAG-tagged proteins from complex mixtures.
- Flow Cytometry: For analyzing the expression of FLAG-tagged proteins on the cell surface.
- Protein Purification: For purifying FLAG-tagged proteins using affinity chromatography .
The monoclonal ANTI-FLAG M2-Peroxidase is a mouse IgG antibody covalently conjugated to horseradish peroxidase (HRP) . This conjugation allows for the detection of FLAG fusion proteins through enzymatic reactions that produce a colorimetric or chemiluminescent signal. The antibody recognizes the FLAG epitope at various positions, including N-terminal, Met-N-terminal, C-terminal, and internal FLAG peptides .
The ANTI-FLAG M2-Peroxidase antibody is typically supplied in a buffered aqueous glycerol solution and is used in various applications such as:
- Immunocytochemistry: Suggested dilution of 1:100 to 1:1000.
- Western Blotting: Suggested dilution of 1:100 to 1:1000.
- ELISA: Suggested dilution of 1:20,000 .
For optimal results, the antibody solution should be diluted in Tris Buffered Saline (TBS) with a pH of 7.4 and 0.15 M NaCl .
