Mouse anti sfpq

Manufactured by Merck Group

Sourced in United States

The Mouse anti-SFPQ is a laboratory reagent used for the detection and analysis of the SFPQ (splicing factor proline and glutamine rich) protein in biological samples. It is a monoclonal antibody that specifically binds to the SFPQ protein, allowing researchers to study the expression, localization, and function of this important cellular component.

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3 protocols using mouse anti sfpq

Kinesin Protein Expression and Detection

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HEK 293T cells or DRG sensory neurons were collected and prepared with lysis buffer (1% NP-40; 50 mM Tris-HCl, pH 7.4; 150 mM NaCl; 2 mM EDTA; protease inhibitor [Sigma-Aldrich]; and phosphatase inhibitor [Life Technologies]). Cell lysates were placed on ice for 20 min and centrifuged at 13,000 rpm for 20 min to collect the supernatant. Lysates were separated with 4–12% Bis-Tris NuPAGE gel (Thermo Fisher Scientific) and blotted with the following primary antibodies: mouse anti-SFPQ (1:1,000; Sigma-Aldrich), rabbit anti-KIF5A (1:2,000; Abcam), rabbit anti-KIF5B (1:2,000; Abcam), rabbit anti-KIF5C (1:2,000; Abcam), rabbit anti-KIF3C (1:500; Abcam), rabbit anti-KIF3A (1:200; Abcam), rabbit anti-KIF1A (1:5,000; Abcam), mouse anti-HAlo (1:1,000), rabbit anti-KLC1 (1:500; Santa Cruz Biotechnology), rabbit anti-KLC2 (1:1,000; Proteintech), rabbit anti-pan actin (1:1,000; Cell Signaling Technologies), mouse anti-Myc (1:500; Santa Cruz Biotechnology), mouse anti-FLAG (1:1,000; Sigma-Aldrich), mouse anti-HA (1:10,000; Thermo Fisher Scientific), and rabbit anti-GFP (1:2m000; Sigma-Aldrich). HRP-conjugated secondary antibodies (1:10,000; BioRad), ECL detection system (VWR International), and SuperSignal West Dura (Thermo Fisher Scientific) were used for chemiluminescent detection.

Fukuda Y., Pazyra-Murphy M.F., Silagi E.S., Tasdemir-Yilmaz O.E., Li Y., Rose L., Yeoh Z.C., Vangos N.E., Geffken E.A., Seo H.S., Adelmant G., Bird G.H., Walensky L.D., Marto J.A., Dhe-Paganon S, & Segal R.A. (2020). Binding and transport of SFPQ-RNA granules by KIF5A/KLC1 motors promotes axon survival. The Journal of Cell Biology, 220(1), e202005051.

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Antibodies for Protein Detection and Localization

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The following antibodies were used: rabbit anti-NONO (Bethyl Laboratories, A300–587A), mouse anti-NONO (BD Biosciences, 611278), rabbit anti-AR (Cell Signaling Technology, 5153S), anti-actin-horseradish peroxidase (HRP; Santa Cruz Biotechnology, sc-47778 HRP), mouse anti-PSPC1 (Sigma-Aldrich, SAB4200503), mouse anti-SFPQ (Sigma-Aldrich, WH0006421M2), mouse anti-FLAG (Sigma-Aldrich, F1804), rabbit anti-fibrillarin (Cell Signaling Technology, 2639S), anti-mouse HRP (Cell Signaling Technology, 7076S), anti-rabbit HRP (Cell Signaling Technology, 7074S), anti-rabbit Alexa 488 (Invitrogen, A11034), anti-mouse Alexa 488 (Invitrogen, A32723), anti-rabbit Alexa 647 (Invitrogen, A32733), anti-mouse Alexa 647 (Invitrogen, A21236) and anti-AR Alexa 488 (Cell Signaling Technology, 7395).

Kathman S.G., Koo S.J., Lindsey G.L., Her H.L., Blue S., Li H., Jaensch S., Remsberg J.R., Ahn K., Yeo G.W., Ghosh B, & Cravatt B.F. (2023). Remodeling oncogenic transcriptomes by small molecules targeting NONO. Nature chemical biology, 19(7), 825-836.

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Lnc-EPAV RNA FISH Assay in BMDMs

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For lnc-EPAV RNA FISH assay, BMDMs cultured on poly-l-lysine-coated coverslips were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, washed, and stored with 70% ethanol at −20°C. Endogenous biotin signal was blocked by using an endogenous biotin blocking kit (Thermo Fisher, USA). BMDMs were incubated with biotin-labeled lnc-EPAV probe at 50°C overnight. The cells were then incubated with Alexa Fluor 488-conjugated avidin (Thermo Fisher, USA) at room temperature for 4 h. For immunofluorescence analysis, cells were sequentially fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, blocked with 5% bovine serum albumin (BSA), and incubated with primary antibody (mouse anti-SFPQ; Sigma, USA) followed by Alexa Fluor 546-conjugated goat anti-mouse secondary antibody (Thermo Fisher, USA). Nuclei were counterstained with DAPI (4',6-diamidino-2-phenylindole). Images were acquired using a Carl Zeiss LSM710 microscope (objective, 40×).

Zhou B., Qi F., Wu F., Nie H., Song Y., Shao L., Han J., Wu Z., Saiyin H., Wei G., Wang P., Ni T, & Qian F. (2019). Endogenous Retrovirus-Derived Long Noncoding RNA Enhances Innate Immune Responses via Derepressing RELA Expression. mBio, 10(4), e00937-19.

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