Anti matrin 3

Manufactured by Fortis Life Sciences

Anti-Matrin-3 is a lab equipment product designed for use in scientific research. It functions to detect and quantify the presence of the Matrin-3 protein in biological samples. This product is intended for research use only and its core function is to provide researchers with a tool to analyze Matrin-3 levels.

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Lab products found in correlation

Anti flag, by Merck Group (2 mentions) Anti hnrnpa1, by Novus Biologicals (1 mentions) Anti caprin 1, by Proteintech (1 mentions) Anti tsg101, by GeneTex (1 mentions) Anti alix, by Santa Cruz Biotechnology (1 mentions) Anti histone h3, by Cell Signaling Technology (1 mentions) Anti gst, by Santa Cruz Biotechnology (1 mentions) Anti cd63, by Santa Cruz Biotechnology (1 mentions) Anti actin, by Santa Cruz Biotechnology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Anti gfp, by Proteintech (1 mentions) Anti hnrnp a1, by Santa Cruz Biotechnology (1 mentions) Anti rbfox2, by Fortis Life Sciences (1 mentions) Anti hnrnp m, by Santa Cruz Biotechnology (1 mentions) Anti hnrnp k, by ABclonal (1 mentions) Anti ddx5, by Abcam (1 mentions) Conformation specific anti rabbit igg, by Cell Signaling Technology (1 mentions) Anti histone h3, by Abcam (1 mentions) Amersham imager 680, by Cytiva (1 mentions)

4 protocols using anti matrin 3

Western Blot Antibody Validation

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The nitrocellulose membranes were blocked and antibodies were applied in 5% milk in TRIS-buffered saline/Tween-20 (TBST, 0.1M TRIS-HCl, 0.9% [w/v] NaCl, 0.1% [v/v] Tween-20, pH 7.5). The antibodies used were anti-NF45/ILF2 (Bethyl, A303-147A-M), anti-DHX9 (Bethyl, A300-855A-M), anti-Matrin-3 (Bethyl, A300-591A-M), anti-hnRNPA1 (Novus, NB100-672), anti-Caprin-1 (Proteintech, 15112-1-AP), anti-GST (Santa Cruz, sc-459), anti-DDX3X (Sigma, HPA001648), anti-actin (Santa Cruz, sc-1616), anti-FLAG (Sigma, A8592), anti-Alix (Santa Cruz, sc-49268), anti-TSG101 (GeneTex, GTX70255), anti-CD63 (Santa Cruz, sc-15363), and anti-Histone H3 (Cell Signaling Technology, 9715). The immunoblotting images were acquired with the Chemidoc MP Imaging System (Bio-Rad) and quantified with the Image Lab software (Bio-Rad).

Kamelgarn M., Chen J., Kuang L., Arenas A., Zhai J., Zhu H, & Gal J. (2016). Proteomic analysis of FUS interacting proteins provides insights into FUS function and its role in ALS. Biochimica et biophysica acta, 1862(10), 2004-2014.

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Western Blot Analysis of Protein Markers

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Protein samples were separated by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) followed by gel transfer to nitrocellulose membrane (Cytiva). The membranes were sequentially incubated with primary antibodies and horseradish peroxidase (HRP)-conjugated secondary antibodies followed by detection with enhanced chemiluminescence system (Millipore) using Amersham Imager 680 (Cytiva). The primary antibodies used in this study were anti-FLAG (Sigma-Aldrich, F3165, 1:8000 diluted), anti-GAPDH (ImB, MM001, 1:3000 diluted), anti-MeCP2 (Diagenode, pAb-052-050), anti-Rbfox1(Millipore, MABE159), anti-Rbfox2 (Bethyl, A300–864A), anti-Rbfox3 (Millipore, MAB377), anti-Matrin3 (Bethyl, A300–591A), anti-hnRNP M (Santa Cruz, sc-20001), anti-hnRNP A1 (Santa Cruz, sc-32301), anti-hnRNP C (Abclonal, A0057), anti-hnRNP H1 (Abclonal, A5924), anti-hnRNP U (Abclonal, A9907), anti-hnRNP F (Abclonal, A5505), anti-NF110 (Abclonal, A2496), anti-hnRNP K (Abclonal, A1701), anti-histone H3 (Abcam, ab1791), anti-DDX5 (Abcam, ab21696), and anti-GFP (Proteintech, 66002-1-Ig). Dilutions of all primary antibodies were 1:1000 if not specifically mentioned. HRP-conjugated secondary antibodies (1:2500 diluted) were anti-mouse IgG (Promega, W4021), anti-rabbit IgG (Promega, W4011), and conformation-specific anti-rabbit IgG (Cell Signaling, 3678).

Jiang Y., Fu X., Zhang Y., Wang S.F., Zhu H., Wang W.K., Zhang L., Wu P., Wong C.C., Li J., Ma J., Guan J.S., Huang Y, & Hui J. (2021). Rett syndrome linked to defects in forming the MeCP2/Rbfox/LASR complex in mouse models. Nature Communications, 12, 5767.

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Western Blot Analysis of Key Signaling Proteins

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Anti-Matrin 3 (part number A300-591A, Bethyl Laboratories, Texas) was used at 1∶2000. The anti-PKA substrate was used at 1∶1000 (part number 9621S, Cell signaling, California), was raised against the motif: RXXps/pS and can recognize phosphorylated substrates of PKA and PKC. Anti-Ro (Santa Cruz, California) and anti-hnrpul1 (part number H00011100-B01, Abnova, California) were used at 1∶1000. Anti 14-3-3γ (Prof A. Aitken, The University of Edinburgh) was used at a dilution of 1∶3000. Secondary antibodies conjugated to IR680 (Invitrogen, Carlsbad) or IR 800 (LI-COR, New Brunswick) dye were used at 1∶15,000 and 1∶5000 respectively and revealed using an Odyssey infrared imager (LI-COR, New Brunswick).

Yamazaki F., Kim H.H., Lau P., Hwang C.K., Iuvone P.M., Klein D, & Clokie S.J. (2014). pY RNA1-s2: A Highly Retina-Enriched Small RNA That Selectively Binds to Matrin 3 (Matr3). PLoS ONE, 9(2), e88217.

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Chromatin IP and Matrin 3 Dependency

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Chromatin IP was performed with the Active Motif ChIP kit (Active Motif, Carlsbad, CA, USA) as directed by the manufacturer. Briefly, 5 × 10⁷ HCT116 cells grown in 15-cm plates were untreated or treated with DOXO (300 nM) for 16 hr. Similarly, PINCR-WT and PINCR-KO cells were untreated or treated with 5-FU (100 µM) for 24 hr. Chromatin was cross-linked with 1% formaldehyde, and cells were lysed and sonicated. Protein–DNA complexes were immunoprecipitated with control IgG or anti-p53 (DO1) (Santa Cruz) or anti-Matrin 3 (Bethyl labs) antibody. The IP material was washed and heated at 65°C overnight to reverse crosslinks. ChIP DNA was column purified (Qiagen) and analyzed by qPCR. Primers flanking the p53 binding sites or the enhancer regions of different genes are listed in Supplementary file 5. To test if association of p53 to the promoter and enhancer regions is Matrin-3-dependent 5 × 10⁷PINCR-WT cells were reverse transfected with CTL siRNAs and two independent Matrin 3 siRNAs. After 48 hr, cells were treated with 5-FU for 24 hr and enrichment of p53 at the promoter and enhancer regions of PINCR targets was determined by ChIP-qPCR as described above.

Chaudhary R., Gryder B., Woods W.S., Subramanian M., Jones M.F., Li X.L., Jenkins L.M., Shabalina S.A., Mo M., Dasso M., Yang Y., Wakefield L.M., Zhu Y., Frier S.M., Moriarity B.S., Prasanth K.V., Perez-Pinera P, & Lal A. (2017). Prosurvival long noncoding RNA PINCR regulates a subset of p53 targets in human colorectal cancer cells by binding to Matrin 3. eLife, 6, e23244.

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