Biolegend, 900801, Anti-HA.11 Epitope Tag Affinity Matrix (Previously Covance catalog# AFC-101P), 1ml

The HA tag (hemagglutinin) is an amino acid sequence derived from the human influenza hemagglutinin surface glycoprotein, corresponding to amino acids 98-106.  It is commonly used as a tag to facilitate detection, isolation, and purification of proteins. The full amino acid sequence is: YPYDVPDYA.
1ml
Verified Reactivity: Influenza, Epitope tag
Reported Reactivity: Species independent
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Monoclonal antibody HA.11 was raised against the twelve amino acid peptide CYPYDVPDYASL.
Formulation: Purified IgG immobilized on Sepharose™ Fast Flow beads (in PBS + 0.03% Thimerosal).
Preparation: The antibody was purified by affinity chromatography.
Storage & Handling: Store between 2-8°C. The matrix may be re-used several times. To strip column after use, wash with several bead volumes of 0.1 M glycine pH 2.8 followed immediately by PBS containing 0.3% Thimerosal or 1mM Sodium Azide as preservative.
Application: Purification, IP
Recommended Usage: Each lot of this antibody is quality control tested by immunoprecipitation. The optimal buffers and matrix concentration should be determined for each specific assay condition. Binding: Tagged protein will bind to matrix in common physiologic buffers with pH in the range of 6.0-7.5, salt from 50-500 mM and in the presence of reasonable levels of detergent. Excess reducing agent should be avoided as the disulfide bridges holding antibody heavy and light chains may be compromised. BioLegend tests Affinity Matrix using an equilibration/binding buffer containing: • 100 mM Tris-HCl (pH 7.5) • 150 mM NaCl • 0.1% Tween 20 • 0.5% BSA • 1 mM beta-mercaptoethanol Washing: After binding, washes with several bead volumes of buffer are recommended. Such buffer may contain increased salt, altered pH, etc. as determined empirically to remove un-tagged proteins. Elution: Several options are available for elution. 1. SDS gel loading buffer may be applied directly to the beads in order to display all bound protein on a polyacrylamide gel/western. Note that gel loading buffer containing reducing agent will also release some antibody heavy and light chains (approx 25 and 50kD, respectively). 2. Competitive elution with epitope peptide. For epitope tag affinity matrices, prepare an elution buffer with epitope tag peptide at 400 ug/mL in 50 mM Tris-HCl (pH 7.5), 50 mM NaCl, 1 mM EDTA (pH 8.0). 3. Chemical Elution. Elution by pH or chaotropic salts is also possible. For elution by pH, either 0.1 M glycine pH 2.8 or 40 mM diethyl-amine pH 11.0 may be used.
Application Notes: This affinity matrix can be used for immunopurification of HA-tagged fusion proteins from crude starting material. On an analytical scale, it can also be used for immunoprecipitating HA-tagged proteins. Monoclonal mouse antibody HA.11 recognizes the peptide epitope, YPYDVPDYA. This second-generation HA antibody is an excellent substitute for the 12CA5 monoclonal antibody. The HA.11 antibody recognizes HA epitopes located in the middle of protein sequences as well as at the N- or C-terminus. HA.11 antibody was purified using protein-G chromatography and was subsequently immobilized onto a Sepharose™ Fast Flow matrix. Sepharose is a trademark of Amersham Biosciences Limited
Application References(PubMed link indicates BioLegend citation): Ferrando A, et al. 2001. Nucleic Acids Res. 29:3685. J Field, et al. 1988. Mol Cell Biol. 8:2159. Bennett BD, et al. 2000. J Biol Chem. 275:37712. (IF, IP, WB) PubMed
Product Citations: Mitxelena J, et al. 2016. Nucleic Acids Res. 44: 5557 - 5570. PubMed Alkan F, et al. 2021. Bioinformatics. 37:2659. PubMed Sivadasan R, et al. 2016. Nat Neurosci. 10.1038/nn.4407. PubMed Karunakaran MM, et al. 2020. Immunity. 52:487. PubMed Gay S, et al. 2018. Mol Cell. 8:e2728. PubMed Bennett B, et al. 2000. J Biol Chem. 275:37712-37717. PubMed Srinivasan B, et al. 2021. PLoS Biol. 19:e3001432. PubMed Dickey D, et al. 2016. J Biol Chem. 291: 11385 - 11393. PubMed
RRID: AB_2564999 (BioLegend Cat. No. 900801)
Biology Area: Cell Biology
Gene ID: NA
UniProt: View information about HA.11 on UniProt.org
Clone: 16B12
Regulatory Status: RUO
Other Names: Hemagglutinin tag, HA tag
Previously: Covance Catalog# AFC-101P
Isotype: Mouse IgG1, κ