Anti-HA tag Antibody, Mouse Monoclonal

RRID Number: AB_2860039

Anti-HA tag Antibody, Mouse Monoclonal の製品情報

製品名
Anti-HA tag Antibody, Mouse Monoclonal
検証済のアプリケーション
WB,ELISA,FCM,ICC/IF,IP
特異性
HA tag
免疫原
A synthetic peptide corresponding to the HA-tag sequence (YPYDVPDYA).
調製
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.
ソース
Monoclonal Mouse IgG1 Clone #10
精製
Protein A
緩衝液
0.2 μm filtered solution in PBS
標識物
Unconjugated
濃度
0.5 μl/Test
状態
Liquid
配送方法
This antibody is shipped as liquid solution at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
保存条件
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 検証済のアプリケーション

アプリケーション 推奨希釈率/用量
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 の画像

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 背景情報

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.
参考文献
  • 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.
  • Infection-derived lipids elicit an immune deficiency circuit in arthropods
    Author
    Shaw, 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;
    Year
    2017
    Journal
    Nat Commun
    Application
    WB/IP

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