HAsignalpeptide The Pierce HA peptide is a crucial reagent in molecular biology, primarily used for the elution of HA-tagged proteins during purification processes. This synthetic peptide, characterized by the sequence NH2-YPYDVPDYA-COOH, mimics a portion of the human influenza hemagglutinin (HA) protein, which is a common epitope tag. Its utility lies in its ability to competitively bind to anti-HA antibodies immobilized on affinity matrices like magnetic beads or agarose, thereby releasing the tagged protein of interest.Pierce Anti-HA Magnetic Beads
Epitope tagging, particularly with the HA tag, has become a standard technique for detecting, purifying, and studying proteins that might not have readily available specific antibodies. The HA tag, a short sequence of nine amino acids (YPYDVPDYA), is fused to a protein of interest. Researchers then utilize reagents like Thermo Scientific Pierce Anti-HA Magnetic Beads or Pierce Anti-HA Agarose to capture these tagged proteins from complex biological samples.
Once the HA-tagged protein is bound to the affinity matrix, the HA peptide serves as the key to its release作者：YO Poluektov·2013·被引用次数：47—Abstract. Processing of antigens for presentation to helper T cells by MHC class II involves HLA-DM (DM) and HLA-DO (DO) accessory molecules.. By introducing a high concentration of the synthetic HA peptide, it outcompetes the HA-tagged protein for binding sites on the anti-HA antibody. This competitive elution allows for the recovery of the purified, untagged protein. Protocols often specify preparing the HA peptide at concentrations such as 1 mg/mL or 2 mg/mL in buffer (e.g.Prepare Pierce HA Peptide at 1mg/mL in TBS. 2. Add one bed volume of 1mg/mL Pierce HA Peptide and incubate for 10-15 minutes at 30°C. Elution may be., TBS) and incubating it with the antibody-bound beads for a short period, typically 5-15 minutes at temperatures ranging from 30°C to 37°C, to achieve effective elution.MHC complexes function as antigenic peptide receptors, collecting peptides inside the cell and transporting them to the cell surface, where the MHC-peptide ...
The primary application of the Pierce HA peptide is in affinity purification techniques, including immunoprecipitation (IP) and co-immunoprecipitation (Co-IP).HA Synthetic Peptide Kits like the Pierce HA-Tag IP/Co-IP Kit are designed to streamline these processes, offering a simple and fast method for studying HA-tagged proteins. Beyond elution, HA peptides can also be used in competitive assays to validate antibody specificity or to block antibody binding.The HA peptide,NH2-YPYDVPDYA-COOH, is a synthetic peptide used to competitively elute fusion proteins containing the influenza hemagglutinin (HA) tag bound ...
While the YPYDVPDYA sequence is standard, variations or related peptides exist for specific research needs. For instance, the DYKDDDDK peptide, recognized by anti-FLAG antibodies, serves a similar elution purpose for FLAG-tagged proteins.HA Synthetic Peptide 5 mg | Buy Online Understanding the specific sequence and its corresponding antibody is critical for successful experimental designActivity-based Crosslinking to Identify Substrates of ... - PMC.
When working with HA-tagged proteins and the HA peptide, several factors warrant consideration:
* Peptide Concentration and Incubation Time: The effectiveness of elution depends on optimizing the concentration of the HA peptide and the duration of incubation. Too little peptide or too short an incubation may result in incomplete elution, while excessive amounts or prolonged incubation could potentially impact protein stabilityActivity-based Crosslinking to Identify Substrates of ... - PMC.
* Purity of the HA Peptide: High-purity synthetic peptides are essential to ensure efficient and specific elution without introducing contaminants. Reputable suppliers like Thermo Scientific provide well-characterized HA synthetic peptidesLife Technologies (Thermo Fisher).Pierce™ Anti-HAMagnetic Beads. 4.65.Pierce™ Anti-HAMagnetic Beads quantity. Add to cart..
* Compatibility with Downstream Assays: Researchers should consider how the eluted protein will be used in subsequent experiments.Activity-based Crosslinking to Identify Substrates of ... - PMC The presence of residual peptide, though usually at low concentrations after elution, might theoretically interfere with certain sensitive downstream applicationsTheHA peptidecomprises the epitope recognized by mouse monoclonal antibody Anti-HA (clone 12CA5), which is commonly used in epitope tagging applications. The ....
* Alternative Elution Methods: While peptide elution is common, other methods like using low pH buffers or chaotropic agents might be employed in specific scenarios, though these can sometimes affect protein integrity or downstream compatibility more significantly.
In summary, the Pierce HA peptide is an indispensable tool for researchers employing HA-tagged proteinsPurified 3xFLAG peptide 3x DYKDDDDK ... - Sigma-Aldrich. Its specific sequence and its role in competitive elution facilitate the isolation and study of target proteins, making it a cornerstone in many molecular biology workflows.HA Peptide - an overview | ScienceDirect Topics
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