Baf155

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom

BAF155 is a laboratory reagent used in research applications. It functions as a component of the SWI/SNF chromatin remodeling complex, which plays a role in regulating gene expression. The specific details and intended use of this product are not provided in order to maintain an unbiased and factual approach.

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Lab products found in correlation

7 protocols using baf155

Western Blot Analysis of Kidney Proteins

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Western blot analysis was performed on stage E13.5 Nf2^UB−/− kidneys or cultured Yap^UB-OE kidneys following doxycycline induction. Unless stated otherwise each western blot lane represents one animal (two kidneys). Kidneys were mechanically homogenized and lysed in RIPA buffer supplemented with proteasome and phosphatase inhibitors. Western blot analysis was performed following standard protocols with the following primary antibodies: CALBINDIN (C9848, Sigma, 1:1,000), BAF155 (SC-9748, SantaCruz Biotechnology, 1:2,000), GAPDH (R9545, Sigma, 1:7,500), HA (11867423001, Roche, 1:1,000), LATS1 (#3477, Cell Signaling Technology, 1:1,000), MST1 (#3682, Cell Signaling Technology, 1:2,000), NF2 (HPA003097, Sigma Prestige Antibodies, 1:2,000), phosho-LATS1 S909 (#9157, Cell Signaling Technology, 1:500), phospho-MST1/2 T183/T180 (#3681, Cell Signaling Technology, 1:2,000), phospho-YAP S127 (#4911, Cell Signaling Technology, 1:2,000) and YAP (sc-101199, SantaCruz Biotechnology, 1:2,000). All uncropped western blots can be found in Supplementary Fig. 8.

Reginensi A., Enderle L., Gregorieff A., Johnson R.L., Wrana J.L, & McNeill H. (2016). A critical role for NF2 and the Hippo pathway in branching morphogenesis. Nature Communications, 7, 12309.

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Western Blot and Gel Filtration Analysis

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Western blots were performed according to standard protocols. The final blots were incubated with fluorescence-conjugated secondary antibodies (Li-Cor Odyssey) and signals were read by the Li-Cor Odyssey system. Superose 6 gel-filtration analyses (Bio-Rad) were performed as previously described (Yan et al., 2008 (link)). The following primary antibodies were used in this study: BRG1-G7, BAF155, BAF170, BAF60A, BAF60B, BAF47, BAF57, and OCT4 (Santa Cruz Biotechnologies); NANOG, H3, and H4 (Cell Signaling Technologies); TUBULIN (Abcam); BAF53A (Bethyl Laboratories); and acetyl-H3 and acetyl-H4 (Millipore).

Zhang X., Li B., Li W., Ma L., Zheng D., Li L., Yang W., Chu M., Chen W., Mailman R.B., Zhu J., Fan G., Archer T.K, & Wang Y. (2014). Transcriptional Repression by the BRG1-SWI/SNF Complex Affects the Pluripotency of Human Embryonic Stem Cells. Stem Cell Reports, 3(3), 460-474.

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Western Blot Analysis of SWI/SNF Proteins

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Total protein was denatured for 10 min at 95 °C, separated on a 4–12% SDS-polyacrylamide gel, and transferred to a PVDF membrane (Immobilon FL, EMD Millipore, Billerica, MA). The membrane was blocked with 5% bovine serum albumin (VWR, Batavia, IL) in PBS containing 0.1% Tween-20 (PBST) for 30 mins at room temperature and then incubated in primary antibodies overnight at 4°C. The primary antibodies used were directed against ARID1A (Santa Cruz Biotechnology Inc., Dallas, TX; sc-32761), PBRM1 (Bethyl Laboratories, Montgomery, TX; A301–591A), BAF155 (Santa Cruz Biotechnology Inc., Dallas, TX; Sc-32763), BAF47 (Santa Cruz Biotechnology Inc., Dallas, TX; Sc-166165), LAMIN B1 (Santa Cruz Biotechnology Inc., Dallas, TX; Sc-377000). The primary antibodies were detected by incubating the membranes in goat-anti-rabbit or goat-anti-mouse secondary antibodies (LI-COR Biotechnology, Lincoln, NE) conjugated to IRDye 800CW or IRDye 680 respectively for 1 h at room temperature, and the signals were visualized using Odyssey Clx imager (LI-COR Biotechnology, Lincoln, NE).

Marian C.A., Stoszko M., Wang L., Leighty M.W., de Crignis E., Maschinot C.A., Gatchalian J., Carter B.C., Chowdhury B., Hargreaves D.C., Duvall J.R., Crabtree G.R., Mahmoudi T, & Dykhuizen E.C. (2018). Small Molecule Targeting of Specific BAF (mSWI/SNF) complexes for HIV Latency Reversal. Cell chemical biology, 25(12), 1443-1455.e14.

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Co-Immunoprecipitation of Chromatin Remodelers

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Co-IP was performed as described in Wu et. al. 2014 (link) (Wu et al., 2014 (link)). Antibodies against Brm (Abcam, ab15597), Brg1 (Santa Cruz, sc10768), V5 (Abcam ab15828), Arid1a (Sigma HPA005456), Arid1b (Santa Cruz, sc32762x), Baf155 (Santa Cruz, sc9746), C/ebpα (Santa Cruz, sc166258), and E2f4 (Milipore 05-312) were used for western blot analysis and/or Co-IP.

Sun X., Chuang J.C., Kanchwala M., Wu L., Celen C., Li L., Liang H., Zhang S., Maples T., Nguyen L.H., Wang S.C., Signer R.A., Sorouri M., Nassour I., Liu X., Xu J., Wu M., Zhao Y., Kuo Y.C., Wang Z., Xing C, & Zhu H. (2016). Suppression of the SWI/SNF component Arid1a promotes mammalian regeneration. Cell stem cell, 18(4), 456-466.

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Chromatin Immunoprecipitation (ChIP) Assays

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Chromatin immunoprecipitation (ChIP) assays were performed as described (20 (link)), using the following antibodies: BAF155 (sc-9746X, Santa Cruz Biotechnology), Brg1 (sc-10768X, Santa Cruz Biotechnology), Brm (sc-28710X, Santa Cruz Biotechnology), RIP140 (ab42126, Abcam, Cambridge, UK, www.abcam.com), H3K9me2 (ab1220, Abcam), H3K4me3 (07-473, Millipore), AcH3 (06-599, Millipore), HP1c (sc-10213, Santa Cruz Biotechnology), Oct4 (2750, cell signaling), Sox2 (sc-17320X, Santa Cruz Biotechnology), c-Jun (sc-1694, Santa Cruz Biotechnology), RAR-α (sc-551, Santa Cruz Biotechnology) and c-Fos (sc-48869, Santa Cruz Biotechnology). Primer used to monitor Oct4 CR1 in ChIP assays was 5′-CCCACAGCTCTGCTCCTCC-3′ (forward) and 5′-GCTCACCTAGGGACGGTTTC-3′ (reverse). Primer used to monitor Nanog CR1 in ChIP assays, was 5′-GGTGGACCCTGCAGGTGGGATTAAC-3′ (forward) and 5′-CCCCTATTCTCCCAGGCACCCAGGC-3′ (reverse).

Wu C.Y., Feng X, & Wei L.N. (2014). Coordinated repressive chromatin-remodeling of Oct4 and Nanog genes in RA-induced differentiation of embryonic stem cells involves RIP140. Nucleic Acids Research, 42(7), 4306-4317.

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Characterizing Chromatin Remodeling Factors

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ARID1A (Sigma-Aldrich #HPA005456, western blot and IHC), ARID1A (Santa Cruz #sc-32761, western blot), ARID1B (Abcam #AB57461, western blot and IHC), ARID2 (Abiocode #R2380–1, western blot), Brg1 (Santa Cruz #sc-10768, western blot), BAF170 (Santa Cruz #sc-17838, western blot), BAF155 (Santa Cruz #sc-9746, western blot), BAF60b (Santa Cruz #sc-101162, western blot), BAF57 (Bethyl #A300–810A, western blot), BAF53 (Santa Cruz #sc-137063, western blot), BAF47 (Santa Cruz #sc-166165, western blot), BAF47 (CST #91735, western blot), BAF45d (Santa Cruz #sc-101106, western blot), Ki-67 (Abcam #AB15580, IHC), HNF4A (CST #3113, IHC), CK19 (Abcam #AB15463, IHC), EpCAM (CST #14452, IHC), Flag (CST #2368, western blot), Ty1 (Diagenode #C15200054, western blot and ChIP).

Wang Z., Chen K., Jia Y., Chuang J.C., Sun X., Lin Y.H., Celen C., Li L., Huang F., Liu X., Castrillon D.H., Wang T, & Zhu H. (2020). Dual ARID1A/ARID1B loss leads to rapid carcinogenesis and disruptive redistribution of BAF complexes. Nature cancer, 1(9), 909-922.

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Immunostaining of Neural Markers

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Antibodies against the following proteins were used: TUJ1 (Covance, MMS425P, 1:500), phospho-histone H3 (Cell Signaling, 9701, 1:200), BrdU (Novus Biologicals, NB500-169, 1:200), p75^NTR (Promega, G3231, 1:100), BRG1 (Santa Cruz, SC-17796 1:200), BAF155 (Santa Cruz, SC-10756, 1:200), BAF170 (Santa Cruz, SC17838, 1:200), and BAF60a (BD Transduction Laboratories, 611728, 1:500). Antibodies from the Developmental Studies Hybridoma Bank were used at a 1:10 dilution including PAX3 and PAX6.

Harmacek L., Watkins-Chow D.E., Chen J., Jones K.L., Pavan W.J., Salbaum J.M, & Niswander L. (2014). A Unique Missense Allele of BAF155, a Core BAF Chromatin Remodeling Complex Protein, Causes Neural Tube Closure Defects in Mice. Developmental neurobiology, 74(5), 483-497.

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