HA Peptideelution protocol The Pierce HA peptide is a crucial reagent in molecular biology, primarily used for the elution of HA-tagged proteins during purification processes.10348. HA SYNTHETIC PEPTIDE 5MG - PIERCE 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.HA-PCNA protein was released from the trap through a two-step competitive elution withHA-peptide. ...HA peptide, hemagglutinin. Size: 1mg. 1mg. 0/ 1mg. In ... 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 interestPierce™ 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.Life Technologies (Thermo Fisher).Pierce™ Anti-HAMagnetic Beads. 4.65.Pierce™ Anti-HAMagnetic Beads quantity. Add to cart.
Once the HA-tagged protein is bound to the affinity matrix, the HA peptide serves as the key to its release. 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.Pierce™ Anti-HA Magnetic Beadsg.HA Synthetic Peptide 5 mg | Buy Online, 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.
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 proteinsPierce Anti-HA Agarose. Beyond elution, HA peptides can also be used in competitive assays to validate antibody specificity or to block antibody binding.
While the YPYDVPDYA sequence is standard, variations or related peptides exist for specific research needsRichard Pierce. For instance, the DYKDDDDK peptide, recognized by anti-FLAG antibodies, serves a similar elution purpose for FLAG-tagged proteins.The attempt to render PRINCIPLES OFPEPTIDESYNTHESIS somewhat resistant to the passing of time could, of course, be only partially successful. Understanding the specific sequence and its corresponding antibody is critical for successful experimental design.HA peptide can be used to competitively elute HA-tagged fusion proteinsfrom immobilized anti-HA magnetic beads. Use HA peptide to elute HA-tagged proteins.
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 stability.
* Purity of the HA Peptide: High-purity synthetic peptides are essential to ensure efficient and specific elution without introducing contaminants.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. Reputable suppliers like Thermo Scientific provide well-characterized HA synthetic peptides.
* Compatibility with Downstream Assays: Researchers should consider how the eluted protein will be used in subsequent experiments作者：AK Sato·2000·被引用次数：103—We tested theHa peptideanalogues for their ability to induce this conformational change using a gel filtration assay. Complexes of each of the peptide .... The presence of residual peptide, though usually at low concentrations after elution, might theoretically interfere with certain sensitive downstream applications.
* 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 proteins. 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.
Join the newsletter to receive news, updates, new products and freebies in your inbox.