Anti-HA tag Antibody, Mouse Monoclonal
RRID Number: AB_2860039
Cat: 100028-MM10
Anti-HA tag Antibody, Mouse Monoclonal General Information
Product name
Anti-HA tag Antibody, Mouse Monoclonal
Validated applications
WB,ELISA,FCM,ICC/IF,IP
Specificity
HA tag
Immunogen
A synthetic peptide corresponding to the HA-tag sequence (YPYDVPDYA).
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.
Source
Monoclonal Mouse IgG1 Clone #10
Purification
Protein A
Formulation
0.2 μm filtered solution in PBS
Conjugate
Unconjugated
Concentration
0.5 μl/Test
Form
Liquid
Shipping
This antibody is shipped as liquid solution at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
This antibody can be stored at 2℃-8℃ for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20℃ to -80℃. Preservative-Free. Avoid repeated freeze-thaw cycles.
Anti-HA tag Antibody, Mouse Monoclonal Validated Applications
| Application | Dilution |
|---|---|
| WB | 1/2000-1/5000 |
| ELISA | 1/2000-1/5000 |
| ICC-IF | 1/200-1/500 |
| IP | 2-8 μL/mg of lysate |
Please Note: Optimal concentrations/dilutions should be determined by the end user.
Anti-HA tag Antibody, Mouse Monoclonal Images
This antibody can be used at 0.5-1 μg/mL with the appropriate secondary reagents to detect HA-tagged protein (amino-terminus HA tag or carboxy-terminus HA tag).
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 (Cat. No. 554714,BD Pharmingen) and subsequently stained with Purified Mouse Anti-HA Tag antibody (Cat No. 100028-MM10) Or positive control antibody for 30 min on ice. Cells were washed twice and incubated with 1 μg of a FITC Goat Anti-Mouse Ig secondary antibody for 30 min on ice. Cells were washed twice and analyzed by flow cytometry.
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℃ 1 hour. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (green).
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℃ 1 hour. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (green) and counterstained with DAPI (blue).
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 μL anti-HA Tag mouse monoclonal antibody and 60 μg of Immunomagnetic beads Protein G.
Primary antibody:
Anti-HA Tag rabbit monoclonal antibody, at 1:100 dilution
Secondary
Goat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.
Developed using the Odyssey technique.
Performed under reducing conditions.
Anti-HA Tag mouse monoclonal antibody at 1:1000 dilution
Lane 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)
Secondary
Goat Anti-Mouse IgG H&L (Dylight800) at 1/15000 dilution.
Developed using the Odyssey technique.
Performed under reducing conditions.
HA tag Background Information
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.
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Infection-derived lipids elicit an immune deficiency circuit in arthropodsAuthorShaw, DK;Wang, X;Brown, LJ;Chávez, AS;Reif, KE;Smith, AA;Scott, AJ;McClure, EE;Boradia, VM;Hammond, HL;Sundberg, EJ;Snyder, GA;Liu, L;DePonte, K;Villar, M;Ueti, MW;de la Fuente, J;Ernst, RK;Pal, U;Fikrig, E;Pedra, JH;Year2017JournalNat CommunApplicationWB/IP
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