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Anti smc3

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Anti-SMC3 is a laboratory antibody product. It recognizes the SMC3 protein, which is a component of the cohesin complex involved in sister chromatid cohesion during cell division.

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Lab products found in correlation

Anti ctcf, by Merck Group (2 mentions) Anti histone h3, by Abcam (2 mentions) Anti ctcf, by Cell Signaling Technology (2 mentions) Anti rad21, by Abcam (2 mentions) Anti plzf, by Santa Cruz Biotechnology (1 mentions) Bovine serum albumin bsa, by ZSGB-BIO (1 mentions) Anti sycp3, by Abcam (1 mentions) Sycp1, by Abcam (1 mentions) Anti vasa, by Abcam (1 mentions) Anti c kit, by Abcam (1 mentions) Anti shp2, by Santa Cruz Biotechnology (1 mentions) Anti dmc1, by Abcam (1 mentions) Vectashield containing 4 6 diamidino 2 phenylindole dapi, by Vector Laboratories (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti v5, by Thermo Fisher Scientific (1 mentions) Anti lamin b1, by Abcam (1 mentions) Anti smc1a, by Fortis Life Sciences (1 mentions) Anti α tubulin, by Santa Cruz Biotechnology (1 mentions) Anti nup153, by Abcam (1 mentions) Anti rpb1 ntd, by Cell Signaling Technology (1 mentions)

6 protocols using anti smc3

1

Comprehensive Protein Detection Protocol

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Anti-CTCF (1:1000; Millipore, 07-729), anti-SMC1A (1:1000: Bethyl, A300-055A), anti-SMC3 (1:1,000; Abcam, ab9263), anti-RAD21 (1:1000; Abcam, ab992), anti-FLAG (1:1000; Sigma-Aldrich, F1804), anti-NUP153 (1:1000; Abcam, ab24700), anti-GAPDH (1:10,000; Sigma-Aldrich, G9545), anti-Histone H3 (1:10,000; Abcam, ab1791), and anti-α-TUBULIN (1:11,000; Santa Cruz, sc-5286) were used in western blot analysis. Note that anti-NUP153 (Abcam, ab24700) can also detect NUP62 and was used to detect NUP62 by western blot analysis. Anti-Rpb1 NTD (3 µl, Cell Signaling, 14958), Anti-CTCF (3 µl, Cell Signaling, 2899S), and anti-SMC3 (3 µg, Abcam, ab9263) were used in ChIP. Anti-LAMIN B1 (1:450; Abcam, ab16048), anti-V5 (1:400; Thermo Fisher Scientific, R960-25), anti-FLAG M2 (1:250; Sigma-Aldrich, F1804), anti-IgG(H+L)-Alexa555 (1:500; Thermo Fisher Scientific, A-21427), and anti-IgG(H+L)-Alexa488 (1:400; Thermo Fisher Scientific, A-11008 and A-32723) were used in immunofluorescence.
Kadota S., Ou J., Shi Y., Lee J.T., Sun J, & Yildirim E. (2020). Nucleoporin 153 links nuclear pore complex to chromatin architecture by mediating CTCF and cohesin binding. Nature Communications, 11, 2606.
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2

Immunofluorescence Staining of Germ Cell Markers

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The prepared sections were blocked with 3% (w/v) bovine serum albumin (BSA; ZSbio) in PBST (0.1% [v/v] Triton X-100 in PBS) for 1 h at room temperature and then incubated with the following primary antibodies overnight at 4°C: anti-Sycp3 (1:200, Abcam, Cambridge, MA, USA), anti-synaptonemal complex protein 1 (Sycp1; 1:200, Abcam), anti-Vasa (1:500, Abcam), anti-c-kit (1:500, Abcam), anti-Shp2 (1:200, Santa Cruz Biotechnology), anti-Plzf (1:500, Santa Cruz Biotechnology), anti-cleaved caspase 3 (1:200, Cell Signaling Technology, Boston, MA, USA), anti-Dmc1 (1:200, Abcam), anti-Smc3 (1:500, Abcam), and anti-DNA repair recombinase rad51 (Rad51; 1:200, Invitrogen). After being washed three times with PBST, the samples were incubated with the following secondary antibodies at a 1:200 dilution for 1 h at 37°C: Alexa Fluor 594/488-labeled anti-rabbit or anti-mouse IgG (YEASEN, Shanghai, China). The slides were subsequently washed three times in PBST and mounted with Vectashield containing 4’-6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA).
Li Y., Liu W.S., Yi J., Kong S.B., Ding J.C., Zhao Y.N., Tian Y.P., Feng G.S., Li C.J., Liu W., Wang H.B, & Lu Z.X. (2019). The role of tyrosine phosphatase Shp2 in spermatogonial differentiation and spermatocyte meiosis. Asian Journal of Andrology, 22(1), 79-87.
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3

Comprehensive Antibody List for Cell Analysis

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The following antibodies were used: anti-CD71-FITC (SouthernBiotech, 1720-02, 0.5 mg ml−1, 1:500)20 (link), anti-SIRT1 (rabbit, Cell Signalling, 2496S, 1:1,000), anti-centromere (Antibodies, 15-234-0001, 1:1,000), anti-rabbit Alexa 488 (Thermo Fisher Scientific, A11034, 1:2,000), goat anti-human Alexa 647 (Thermo Fisher Scientific, A21445, 1:2,000), anti-DSCC1 (H0079075-B01P, Novus Biologicals, 1:1,000), anti-HSP90 (F-8, Santa Cruz, 1:10,000), anti-HP1γ (05-690, Millipore, 1:1,000), goat-anti-mouse-PO (DAKO, P044701, 1:2,000), anti-SMC3 (Abcam, AB 9263, 1:250), anti-SMC3 (Thermo Fisher Scientific, A300-060A, 1:1,000), anti-acetyl SMC3 mouse (Sigma-Aldrich, MABE1073, 21A7, Lys105/106, 385016, 1:1,000), anti-p53 (Cell Signaling Technology, 1C12, 2524S), anti-acetyl p53 (p53-K382Ac, Abcam, ab75754, EPR358(2) to p53 acetyl K382, 1:1,000), anti-phosphorylated-histone H2A.X (Ser139) (JBW301, Sigma-Aldrich, 05-636-I, 1:1,000), anti-β-actin (Merck, A5441, 1:10,000, 5% milk), anti-GAPDH (6C5, Abcam, ab8245, 1:1,000), anti-p21 (Abcam, ab109520, 1:1,000). Uncropped western blots are provided in the Supplementary Information.
Adams D.J., Barlas B., McIntyre R.E., Salguero I., van der Weyden L., Barros A., Vicente J.R., Karimpour N., Haider A., Ranzani M., Turner G., Thompson N.A., Harle V., Olvera-León R., Robles-Espinoza C.D., Speak A.O., Geisler N., Weninger W.J., Geyer S.H., Hewinson J., Karp N.A., Fu B., Yang F., Kozik Z., Choudhary J., Yu L., van Ruiten M.S., Rowland B.D., Lelliott C.J., del Castillo Velasco-Herrera M., Verstraten R., Bruckner L., Henssen A.G., Rooimans M.A., de Lange J., Mohun T.J., Arends M.J., Kentistou K.A., Coelho P.A., Zhao Y., Zecchini H., Perry J.R., Jackson S.P, & Balmus G. (2024). Genetic determinants of micronucleus formation in vivo. Nature, 627(8002), 130-136.
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4

Western Blot Analysis of Rab27a and SMC3

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Western blot was performed as described previously [13 (link)]. 30 µg of protein derived from cell lines was used. Primary antibody incubation was performed overnight at 4 ºC. Antibody dilutions used were: anti-Rab27a 1:500 (Abnova Cat# H00005873-M02, RRID:AB_519010), anti-SMC3 1:1000 (Abcam Cat# ab9263, RRID:AB_307122). After washing steps, membranes were incubated with the respective HRP-conjugated secondary antibody at 1:5000 dilution for 1 h at RT (Anti-mouse IgG HRP-linked antibody Advansta Cat# R-05071–500, RRID:AB_10718209; Anti-rabbit IgG HRP-linked antibody Cell Signaling Technology Cat# 7074, RRID:AB_2099233). β-actin (Sigma-Aldrich Cat# A3854, RRID:AB_262011) was used for loading control.
Bastos N., Castaldo S.A., Adem B., Machado J.C., Melo C.A, & Melo S.A. (2023). SMC3 epigenetic silencing regulates Rab27a expression and drives pancreatic cancer progression. Journal of Translational Medicine, 21, 578.
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5

ChIP and Immunoblot Antibody Panel

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The following antibodies were used in ChIPs and/or immunoblots: anti-histone H3 (Abcam ab1791), anti-H3K27me3 (Active Motif #39155), anti-H3K4me3 (Active Motif #39159), anti-EZH2 (Active Motif #39875), anti-RING1B (Abcam ab3832), anti-LANA (Advanced Biotechnologies #13-210-100), anti-CTCF (Millipore, #07-729), anti-RAD21 (Abcam, ab992), anti-SMC3 (Abcam, ab9263), anti-NIPBL (Bethyl Laboratories, A301-779A), anti-K8 (Abcam ab36617), anti-RNA polymerase II (RNAPII) (Abcam), and anti-actin (Abcam). Anti-K3 and anti-RTA antibodies were generous gifts from Drs. Jae U. Jung (University of Southern California) and Yoshihiro Izumiya (University of California, Davis).
Toth Z., Smindak R, & Papp B. (2017). Inhibition of the lytic cycle of Kaposi’s sarcoma-associated herpesvirus by cohesin factors following de novo infection. Virology, 512, 25-33.
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6

ChIP-seq Profiling of Histone Marks and Transcription Factors

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Chromatin immunoprecipitation (ChIP) was performed as previously described47 (link),48 for the following antibodies: anti-H3K27ac (Hiroshi Kimura Laboratory, clone CMA30949 (link), 10 μg/20 M cells), anti-H3K27me3 (Hiroshi Kimura Laboratory, clone CMA32349 (link), 5 μg/10 M cells), anti-CTCF (Cell Signaling Technology, 3418, clone D31H12, lot #1, 10 μL/20 M cells), anti-RAD21 (Katsuhiko Shirahige Laboratory50 (link), 10 μg/20 M cells) and anti-SMC3 (Abcam ab9263, lot # GR290533-17 and GR3221084-8, 10 μg/20 M cells), anti-Pol II (Hiroshi Kimura Laboratory, clone C13B949 (link), 5 μg/10 M cells). Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs, E7645L) according to the manufacturer’s instructions except that size selection was performed after PCR amplification using AMPure XP beads (Beckman Coulter, A63881). Samples were sequenced paired-end using 50 bp reads on the Illumina platforms.
Olan I., Parry A.J., Schoenfelder S., Narita M., Ito Y., Chan A.S., Slater G.S., Bihary D., Bando M., Shirahige K., Kimura H., Samarajiwa S.A., Fraser P, & Narita M. (2020). Transcription-dependent cohesin repositioning rewires chromatin loops in cellular senescence. Nature Communications, 11, 6049.
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