Mouse anti-HA tag HA Monoclonal Antibody | anti-HA antibody

Anti-HA tag Antibody, Mouse Monoclonal

Cat. Num. AAA8100024

• Reactivity: HA tag
• Applications: Western Blot, ELISA, Flow Cytometry, Functional Assay, Immunocytochemistry, Immunofluorescence, Immunoprecipitation
• Purity: Protein A
• Synonyms: HA; Monoclonal Antibody; Anti-HA tag Antibody; Mouse Monoclonal; Anti-HA Tag Antibody; Mouse MAb; anti-HA antibody

• Host: Mouse
• Reactivity: HA tag
• Clonality: Monoclonal
• Isotype: Mouse IgG1
• Clone Number: 10
• Specificity: Recognize N-terminal and C-terminal HA Tag in fusion proteins.
• Purity/Purification: Protein A
• Form/Format: Liquid; 0.2um filtered solution in PBS
• Concentration: 0.5ul/Test (varies by lot)

• Applicable Applications for anti-HA antibody: Western Blot (WB), ELISA (EIA), Flow Cytometry (FC/FACS/FCM), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP)
• Application Notes: WB: 1:2000-1:5000; ELISA: 1:2000-1:5000; ICC/IF: 1:200-1:500; IP: 2-8uL/mg of lysate
• Immunogen: A synthetic peptide corresponding to the HA-tag sequence (YPYDVPDYA)
• Conjugation: Unconjugated
• Preparation: This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, a synthetic peptide corresponding to the HA-tag sequence. The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography.
• Preparation and Storage: This antibody can be stored at 2-8 degree C for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20 degree C to -80 degree C. Preservative-Free. Avoid repeated freeze-thaw cycles.

Testing Data

(This antibody can be used at 0.5-1 ug/mL with the appropriate secondary reagents to detect HA-tagged protein (amino-terminus HA tag or carboxy-terminus HA tag).)

Flow Cytometry (FC/FACS)

(Profile of anti-HA Tag reactivity on 293 transfected cells analyzed by flow cytometry. 293 cells transfected with HA-ARG1 (Figure A), HA-mFABP4 (Figure B), ARG1-HA (Figure C) and mFABP4-HA (Figure D) were fixed and permeabilized according to the manufacturer's manual and subsequently stained with Purified Mouse Anti-HA Tag antibody Or positive control antibody  for 30 min on ice. Cells were washed twice and incubated with 1 ug of a FITC Goat Anti-Mouse Ig secondary antibody for 30 min on ice. Cells were washed twice and analyzed by flow cytometry.)

Immunofluorescence (IF)

(Immunofluorescence staining of HA-Tag in 293 cells, transfected with HA-ARG1 (L) or mock-transfected (R). Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with SBI Mouse anti-HA-tag monoclonal antibody at 37 degree C 1 hour. Then cells were stained with the Alexa Fluor 488-conjugated Goat Anti-mouse IgG secondary antibody (green).)

Immunofluorescence (IF)

(Immunofluorescence staining of HA-Tag in 293 cells, transfected with HA-ARG1 (Figure A), HA-mFABP4 (Figure B), ARG1-HA (Figure C) and mFABP4-HA (Figure D). Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with Mouse anti-HA-Tag monoclonal antibody at 37 degree C 1 hour. Then cells were stained with the Alexa Fluor 488-conjugated Goat Anti-mouse IgG secondary antibody (green) and counterstained with DAPI (blue).)

Immunoprecipitation (IP)

(Anti-HA Tag was immunoprecipitated using:Lane A: HA-ARG1-myc transfected 293 cell lysate (0.5mg)Lane B: myc-ARG1-HA transfected 293 cell lysate (0.5mg)2 uL anti-HA Tag mouse monoclonal antibody and 60 ug of Immunomagnetic beads Protein G.Primary antibody:Anti-HA Tag rabbit monoclonal antibody, at 1:100 dilutionSecondaryGoat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.Developed using the Odyssey technique.Performed under reducing conditions.)

Western Blot (WB)

(Anti-HA Tag mouse monoclonal antibody at 1:1000 dilutionLane A: HA-GST (Recombinant protein)(30ng)Lane B: GST-HA (Recombinant protein)(10ng)Lane C: HA-ARG1-myc transfected 293 cell lysate (2ug)Lane D: HA-mFABP4-myc transfected 293 cell lysate (2ug)Lane E: myc-mFABP4-HA transfected 293 cell lysate (0.5ug)Lane F: myc-ARG1-HA transfected 293 cell lysate (2ug)SecondaryGoat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.Developed using the Odyssey technique. Performed under reducing conditions.)

Related Product Information for anti-HA antibody

The human influenza hemagglutinin (HA) epitope was derived from the protein on the HA virus. HA tag is the amino acid sequence of a YPYDVPDYA tag and a particularly common tags. HA is tagged onto proteins for study and analysis and often used in fusion protein-related research through the realization of detection of HA-tag for the detection of fusion proteins. Antibodies to HA-tag are the key of the most important tools in the detection and often used in purification of HA tag-related proteins, study of the protein interaction and detection of target protein expression.

References

Zhao X, et al. (2013) Several affinity tags commonly used in chromatographic purification. J Anal Methods Chem 2013 581093.Monti M, et al. (2007) Functional proteomics: Protein-protein interactions in vivo. Ital J Biochem 56 (4): 310-314.Brothers SP, et al. (2003) Unexpected effects of epitope and chimeric tags on gonadotropin-releasing hormone receptors: Implications for understanding the molecular etiology of hypogonadotropic hypogonadism. J Clin Endocrinol Metab 88 (12): 6107-6112.