Smc1a

Manufactured by Fortis Life Sciences

The SMC1A is a lab equipment product that serves as a core structural maintenance of chromosomes protein. It plays a crucial role in sister chromatid cohesion and chromosome segregation during cell division.

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Lab products found in correlation

Nipbl, by Fortis Life Sciences (2 mentions) Goat anti rabbit po, by Agilent Technologies (2 mentions) Hsp90, by Santa Cruz Biotechnology (1 mentions) Goat anti mouse po, by Agilent Technologies (1 mentions) Igg rabbit, by Merck Group (1 mentions) V9131, by Merck Group (1 mentions) Hnf4a, by Santa Cruz Biotechnology (1 mentions) Vinculin, by Merck Group (1 mentions) Foxa1, by Santa Cruz Biotechnology (1 mentions) Fosl2, by Santa Cruz Biotechnology (1 mentions) Na9340, by GE Healthcare (1 mentions) Na931, by GE Healthcare (1 mentions) Nupage novex 4 12 bis tris protein gel, by Thermo Fisher Scientific (1 mentions) H 210, by Santa Cruz Biotechnology (1 mentions) Nupage mops buffer, by Thermo Fisher Scientific (1 mentions) Rad21, by Santa Cruz Biotechnology (1 mentions) Stag2, by Santa Cruz Biotechnology (1 mentions) Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions) Immobilon p, by Merck Group (1 mentions) Rad21, by Abcam (1 mentions)

Spelling variants of this product

Anti-SMC1A (4 citations) SMC1A antibody (2 citations)

5 protocols using smc1a

Antibody Validation for Protein Analysis

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Western Blots were performed using the following antibodies: WAPL (Santa Cruz, A-7), SCC4 (Abcam, ab46906), SCC2 N-terminal (Santa Cruz, C-9), SCC2 C-terminal (Absea, serum of KT55), HSP90 (Santa Cruz, H-114), CDK4 (Santa Cruz, C-22) and Actin (Santa Cruz, I-19). All antibodies were used at 1:1000 dilution. Secondary antibodies Goat anti-Rabbit-PO, Goat anti-Mouse-PO, Rabbit anti-Goat-PO (DAKO) and Rabbit anti-Rat-PO (Santa Cruz, sc2006) were used at 1:2000 dilution. For immunofluorescence we used the SCC1 (Millipore, 05-908) antibody in a 1:100 dilution. CTCF (Millipore, 07-72), SMC1A (Bethyl, A300-055a) and IgG Rabbit (Sigma, I5006) were used for chromatin immuno-precipitations.

Haarhuis J.H., van der Weide R.H., Blomen V.A., Yáñez-Cuna J.O., Amendola M., van Ruiten M.S., Krijger P.H., Teunissen H., Medema R.H., van Steensel B., Brummelkamp T.R., de Wit E, & Rowland B.D. (2017). The Cohesin Release Factor WAPL Restricts Chromatin Loop Extension. Cell, 169(4), 693-707.e14.

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Lentiviral Knockdown of Transcription Factors

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Plasmid catalog numbers and shRNA sequences are provided in Supplementary Table S3. Lentiviruses were produced as previously described58 (link). Cells were transduced with the lentiviruses for 24 h before puromycin selection for the indicated time. Validation of efficiency were performed by Western blot (Supplementary Figs 1 and 3) using MED1 (Bethyl A300-793A, 1:5000), SMC1A (Bethyl, A300-055A, 1:5000), NIPBL, (Bethyl, A301-779A, 1:1000), ERa (Santa Cruz, sc-543x, 1:5000), FOXA1 (Santa Cruz, sc-101058 1:1000), FOXA2 (Abnova, 89-019-034, 1:1000), HNF4a (Santa Cruz, sc-8987x, 1:1000), FOSL2 (Santa Cruz, sc-604x, 1:1000), GAPDH (Pierce, PIMA515738, 1:25000) and Vinculin (SIGMA, V9131, 1:50000).

Fournier M., Bourriquen G., Lamaze F.C., Côté M.C., Fournier É., Joly-Beauparlant C., Caron V., Gobeil S., Droit A, & Bilodeau S. (2016). FOXA and master transcription factors recruit Mediator and Cohesin to the core transcriptional regulatory circuitry of cancer cells. Scientific Reports, 6, 34962.

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Analyzing Cohesin Complex Protein Levels

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Protein was extracted from equal cell numbers in a high urea buffer (48% urea, 15 mM Tris-HCl, pH 7.5, 8.7% glycerin, 1% SDS, 0.004% bromophenol blue, and 143 mM β-mercaptoethanol). Protein levels were measured using the Bradford assay. Equal amounts of protein were loaded on NuPAGE Novex 4–12% Bis-Tris protein gel (NP0321; Life Technologies) and run with NuPAGE MOPS buffer (NP0001; Life Technologies). Proteins were transferred to Immobilon P (IPVH00010; EMD Millipore). Membrane was blocked in 5% nonfat milk in TBST and incubated with antibodies for Rad21 (H-210; Santa Cruz Biotechnology, Inc.), Smc1a (A300-055A; Bethyl Laboratories, Inc.), Smc3 (ab9263; Abcam), Stag2 (J-12; Santa Cruz Biotechnology, Inc.) and Actin B (C4; EMD Millipore). Secondary antibodies used were HRP conjugated (NA9340 and NA931; GE Healthcare). Blot was visualized using ECL (PI34077 and PI34095; Thermo Fisher Scientific).

Mullenders J., Aranda-Orgilles B., Lhoumaud P., Keller M., Pae J., Wang K., Kayembe C., Rocha P.P., Raviram R., Gong Y., Premsrirut P.K., Tsirigos A., Bonneau R., Skok J.A., Cimmino L., Hoehn D, & Aifantis I. (2015). Cohesin loss alters adult hematopoietic stem cell homeostasis, leading to myeloproliferative neoplasms. The Journal of Experimental Medicine, 212(11), 1833-1850.

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Antibody Characterization for Chromatin Study

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The following are the antibodies used in this study: NIPBL (Bethyl Laboratories, A301-779A; Thermo Fisher Scientific, MA1-72534), YY1 (Active Motif, 61779), H3K27ac (Active Motif, 39133), GR (Santa Cruz Biotechnology, sc-393232; Thermo Fisher Scientific, MA1-510), SMC1a (Bethyl Laboratories, A300-055A; Abcam, ab133643), SMC3 (Abcam, ab9263), RAD21 (Abcam, ab992), H2B (Abcam, ab61250), and tubulin (Abcam, ab6160).

Rinaldi L., Fettweis G., Kim S., Garcia D.A., Fujiwara S., Johnson T.A., Tettey T.T., Ozbun L., Pegoraro G., Puglia M., Blagoev B., Upadhyaya A., Stavreva D.A, & Hager G.L. (2022). The glucocorticoid receptor associates with the cohesin loader NIPBL to promote long-range gene regulation. Science Advances, 8(13), eabj8360.

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Immunoblotting Analysis of DNA Damage Response Pathways

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The following antibodies were used: Phospho-ATM/ATR Substrate Motif (pS/TQ) (1:1,000; Cell Signaling, #6966), NIPBL (1:1,000; Santa Cruz, sc-374625), WAPL (1:1,000; Cell Signaling, 77428S), PDS5A (1:1,000; Bethyl A300–089A), PDS5B/APRIN (1:1,000; Novus, NB100–755), Flag (1:1,000; Sigma, F1804), SMC1A (1:1,000, Bethyl A300–055A), pSMC1A S957 (1:1,000 [WB], 1:200 [IF]; Cell Signaling, 4805S), 53BP1 (1:200 [IF], Bethyl, A300–237A); γH2AX (1:200 (IF); Millipore 05–636), ATM (1:1,000; Sigma, A1106), pATM S1981 (1:1,000, Rockland, 200–301–400), CHK2 (1:1,000, Cell Signaling, 2662), pCHK2 T68 (1:1,000; Cell Signaling, 2661), DNAPK (1:1,000; Abcam, ab70250), pDNAPK S2056 (1:1,000; Abcam, ab18192), CHK1 (1:1,000, Epitomics, 2865–1), pCHK1 S345 (1:1,000, Epitomics, S0660). For immunoblotting proteins were transferred to nitrocellulose after separation on an acrylamide gel, blocked in 5% milk in TBS/tween. Primary antibodies were diluted at the indicated dilutions above in 5% bovine serum albumin (BSA) in TBS/t. Proteins were visualized by staining with Alexfluor 700 or 800 conjugated secondary antibodies on an Odyssey CLx imaging system.

Bass T.E., Fleenor D.E., Burrell P.E, & Kastan M.B. (2023). ATM REGULATION OF THE COHESIN COMPLEX IS REQUIRED FOR REPRESSION OF DNA REPLICATION AND TRANSCRIPTION IN THE VICINITY OF DNA DOUBLE STRAND BREAKS. Molecular cancer research : MCR, 21(3), 261-273.

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