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Fitc conjugated anti ha monoclonal antibody

Manufactured by Merck Group

The (FITC)-conjugated anti-HA monoclonal antibody is a laboratory reagent used for the detection and identification of proteins tagged with the HA (hemagglutinin) epitope. The antibody is conjugated with the fluorescent dye FITC (fluorescein isothiocyanate), allowing for visualization and analysis of HA-tagged proteins using techniques such as immunoblotting, immunoprecipitation, and flow cytometry.

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2 protocols using fitc conjugated anti ha monoclonal antibody

1

Immunofluorescence Staining of Flagellar Proteins

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Cells were allowed to adhere to the coverslips were fixed with cold methanol (−20°C), rehydrated with phosphate-buffered saline (PBS) and then blocked in 3% bovine serum albumin in PBS. Immunostaining was performed by incubating the fixed cells with the primary antibody for 1 h at room temperature. The following primary antibodies were used: fluorescein isothiocyanate (FITC)-conjugated anti-HA monoclonal antibody (1:400 dilution; Sigma-Aldrich), L8C4 (anti-PFR2 monoclonal antibody [MAb]; 1:50 dilution) (31 (link)), anti-TbSAS-6 polyclonal antibody (1:400 dilution) (11 (link)), anti-TbBLD10 polyclonal antibody (1:400 dilution), and YL 1/2 (1:1,000 dilution; Millipore). The following secondary antibodies were used: Cy3-conjugated anti-mouse IgG (1:400 dilution; Sigma-Aldrich), FITC-conjugated anti-rat IgG (1:400 dilution; Sigma-Aldrich), and Cy3-conjugated anti-rabbit IgG (1:400 dilution; Sigma-Aldrich). Cells were washed with PBS, mounted with Vectashield mounting medium containing 4′,6-diamidino-2-phenylindole (Vector Labs), and imaged with an inverted fluorescence microscope (Olympus IX71) equipped with a cooled charge-coupled-device (CCD) camera (model Orca-ER; Hamamatsu) and a PlanApo N 60×, 1.42-numeric aperture differential inference contrast objective. Images were acquired using the Slidebook software (version 5; Intelligent Imaging Innovations).
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2

Immunostaining of HA-tagged Proteins

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Cells were washed once with PBS, settled onto glass coverslips for 30 min at room temperature, and fixed with cold methanol for 30 min at − 20 °C. Cells were rehydrated with PBS and incubated with the blocking buffer (3% BSA in PBS) for 1 h at room temperature. Cells were incubated with the FITC-conjugated anti-HA monoclonal antibody (1:400 dilution, Sigma Aldrich, Clone HA7) for 1 h at room temperature and washed three times with PBS. Coverslips were washed three times with PBS and mounted with DAPI-containing VectaShield mounting medium (Vector Labs). Immunostained cells were imaged with an inverted fluorescence microscope (Olympus IX71) equipped with a cooled CCD camera (model Orca-ER, Hamamatsu) and a PlanApo N 60 × 1.42-NA lens. Images were acquired using the Slidebook5 software and processed using the Adobe Photoshop software.
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