Anti srpk1 monoclonal antibody

Manufactured by BD

The Anti-SRPK1 monoclonal antibody is a laboratory reagent used for the detection and quantification of SRPK1 protein in various biological samples. It is a highly specific and sensitive tool for researchers studying the role of SRPK1 in cellular processes and disease mechanisms.

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3 protocols using anti srpk1 monoclonal antibody

Protein Extraction and Western Blot Analysis

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Cells were lysed in 200 μl of 1% Triton buffer (1% Triton X-100, 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, and 1 mM PMSF). Whole cell extracts were clarified by centrifugation for 15 min at 13,000 g in a microcentrifuge, and analyzed on 10% SDS-PAGE. For non-reducing conditions, DTT (90 mM) was omitted from the protein loading buffer. Western blotting was performed with an anti-SRPK1 monoclonal antibody (BD Biosciences) or the M5 anti-FLAG monoclonal antibody (Sigma), an alkaline phosphatase-coupled goat anti-mouse secondary antibody, and 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium substrate. Gel loading was adjusted to give equivalent cell numbers in each lane. Coomassie Brilliant Blue (CBB) staining of bacterially produced recombinant proteins was performed according to standard procedures.

Koutroumani M., Papadopoulos G.E., Vlassi M., Nikolakaki E, & Giannakouros T. (2017). Evidence for disulfide bonds in SR Protein Kinase 1 (SRPK1) that are required for activity and nuclear localization. PLoS ONE, 12(2), e0171328.

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Comprehensive Biochemical Reagents Protocol

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ATP, Mops, Tris, MgCl₂, NaCl, EDTA, glycerol, sucrose, acetic acid, lysozyme, DNAse, RNAse, Phenix imaging film, BSA, Protein G–agarose, Ni-resin and liquid scintillant were obtained from Fisher Scientific. ³²P-ATP was obtained from NEN Products. RNA (AGGCGGAGGAAGC) was purchased from Integrated DNA Technologies. siRNA for CLK1 was obtained from Bioneer. Protease inhibitor cocktail was obtained from Roche and TG003 was obtained from Sigma. Anti-CLK1 monoclonal antibody was purchased from Aviva Systems Biology and anti-SRPK1 monoclonal antibody was purchased from BD Biosciences. InstantBlue was purchased from Expedeon, Hybond ECL nitrocellulose blotting membrane was purchased from Amersham and the KinaseMax™ Kit was purchased from Ambion.

Aubol B.E., Wu G., Keshwani M.M., Movassat M., Fattet L., Hertel K.J., Fu X.D, & Adams J.A. (2016). Release of SR Proteins from CLK1 by SRPK1: A Symbiotic Kinase System for Phosphorylation Control of Pre-mRNA Splicing. Molecular cell, 63(2), 218-228.

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Reagents for Protein Purification and Analysis

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ATP, Mops, HEPES, Tris, MgCl₂, MnCl_2, NaCl, EDTA, Brij 35, glycerol, acetic acid, lysozyme, DNAse, RNAse, Phenix imaging film, BSA, Ni-resin and liquid scintillant were obtained from Fisher Scientific. γ-³²P-ATP was obtained from NEN Products. SRPIN340 was obtained from Sigma. FuGene reagent was obtained from Promeg and Lipofectamine 2000 was obtained from ThermoFisher. Protease inhibitor cocktail, EGF, and TG003 were obtained from Roche. Anti-SRPK1 monoclonal antibody was purchased from BD Biosciences. Anti-CLK1 polyclonal antibody was purchased from Aviva. Anti-SRSF1 monoclonal antibody was purchased from Life Tech. Anti-GFP monoclonal antibody, anti-HA monoclonal antibody, anti-His monoclonal antibody and Protein G beads were purchased from Cell Signaling. Anti-GST monoclonal antibody was purchased from BioLegend. InstantBlue was purchased from Expedeon.

Aubol B.E., Keshwani M.M., Fattet L, & Adams J.A. (2018). Mobilization of A Splicing Factor Through A Nuclear Kinase-Kinase Complex. The Biochemical journal, 475(3), 677-690.

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