HeLa cells transfected with plasmids encoding GFP-TAF15-RGG were grown on glass coverslips for 48 h. After the incubation period, the cell coverslips were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) for 20 min at room temperature, and excess aldehyde was quenched with 100 mM Tris-HCl pH 7.5. Cells were then permeabilized with 0.2% Triton X-100 in PBS for 10 min and blocked with 0.5% fish skin gelatin (FSG) in PBS for 30 min. Probing with the primary (goat polyclonal anti-GFP diluted 1:1000, SICGEN Antibodies; mouse monoclonal anti-SRPK1 diluted 1:150, BD Biosciences, San Jose, CA, USA) and secondary (Alexa 488 donkey anti-goat diluted 1:350, Abcam; Alexa 488 donkey anti-mouse diluted 1:400, Invitrogen; TRITC-conjugated goat anti-mouse, diluted 1:800, Molecular Probes) and DNA staining (propidium iodide) were performed as previously described [18 (link)]. After three washes, the coverslips were mounted with a mounting medium (0.01% p-phenylenediamine and 50% glycerol in PBS) and visualized with a Nikon confocal microscope using the EZ-C1 3.20 software.
Mouse monoclonal anti srpk1
Manufactured by BD
Sourced in United States
Mouse monoclonal anti-SRPK1 is a laboratory reagent that specifically binds to the SRPK1 protein. SRPK1 is a serine/threonine-protein kinase involved in the regulation of pre-mRNA splicing. This product can be used for detection and quantification of SRPK1 in various experimental applications.
Automatically generated - may contain errors
2 protocols using mouse monoclonal anti srpk1
1
Visualization of GFP-TAF15-RGG in HeLa cells
2
Immunofluorescence Staining of SRSF1 and SRPK1
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For immunofluorescence, cells were grown to 80% confluence on glass cover slips. After treatment the cells were washed with PBS, fixed for 5 min with 4% PFA and then washed with PBS in 0.05% Triton X (PBS-TritonX). The cells were blocked in 5% normal goat serum in PBS-TritonX for 1 h, washed thrice with PBS-TritonX and incubated overnight with 2 μg ml−1 of mouse monoclonal SRSF1 (96; sc-33652, Santa Cruz) or 2 mg ml−1 mouse monoclonal anti-SRPK1 (BD Biosciences, Franklin Lakes, NJ, USA, 611072). The cells were washed thrice with PBS-Triton and incubated with goat anti-mouse Alexa Fluor 555 diluted 1 : 100 in 1 × PBS for visualisation and counterstained for the nucleus with Hoescht. Images were taken at × 40 objective using a microscope (Nikon Eclipse E400, Nikon, Surrey, UK).
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