Cyclin b1 gns1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Cyclin B1 (GNS1) is a protein that plays a crucial role in the regulation of the cell cycle. It is an essential component of the mitosis-promoting factor, which drives the progression of cells through the G2/M phase transition. Cyclin B1 (GNS1) functions as a regulatory subunit of the cyclin-dependent kinase 1 (CDK1) enzyme, activating it and promoting the initiation of mitosis.

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Lab products found in correlation

β actin, by Merck Group (2 mentions) Alexa fluor 488 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (1 mentions) Propidium iodide, by Merck Group (1 mentions) Phosphatase inhibitor cocktail 2, by Merck Group (1 mentions) Nocodazole, by Merck Group (1 mentions) Protease inhibitor cocktail 1, by Merck Group (1 mentions) 5 bromo 2 deoxyuridine, by Merck Group (1 mentions) Myc 9e10, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488 conjugated goat anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions) Hoechst 33342, by Bio-Rad (1 mentions) Cyclin a c 19, by Santa Cruz Biotechnology (1 mentions) β actin ac 15, by Santa Cruz Biotechnology (1 mentions) Mops buffer, by Thermo Fisher Scientific (1 mentions) Jbw301, by Merck Group (1 mentions) Atr n 19, by Santa Cruz Biotechnology (1 mentions) Btf 608a, by Fortis Life Sciences (1 mentions) Atm 2c1, by Santa Cruz Biotechnology (1 mentions) Patr s428, by Cell Signaling Technology (1 mentions) Brca2 h 300, by Santa Cruz Biotechnology (1 mentions)

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Cyclin B1 (162 citations)

5 protocols using cyclin b1 gns1

Cell Cycle Regulation Assay Protocol

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Propidium iodide, nocodazole (#M1404), SALL2 (#HPA004162) polyclonal antibody, protease inhibitor cocktail I (# P8340), phosphatase inhibitor cocktail II (P5726), and 5‐bromo‐2′‐deoxyuridine (# B5002) were purchased from Sigma‐Aldrich Chemicals (St. Louis, MO, USA). SALL2 antibody used for ChIP experiments was obtained from Bethyl Lab (Montgomery, TX, USA). Cyclin A (C‐19, #SC‐596) polyclonal antibody and cyclin B1 (GNS1, #SC‐245), cyclin D1 (DCS‐6, #SC‐20044), cyclin E1 (E‐4, #SC‐377100), p21 (F‐5, #6246), Myc (9E10, #SC‐40), and β‐actin (AC‐15, #SC‐69879) monoclonal antibodies were obtained from Santa Cruz Biotechnology (San Diego, CA, USA). The SV40 large T antigen expression pBSSVD2005 plasmid was a gift from David Ron (Addgene plasmid # 21826), the plasmid containing the CCNE1 promoter was a gift from Bob Weinberg (Addgene plasmid # 8458) (Geng et al., 1996), and the CCND1 promoter pGL3Basic was a gift from Frank McCormick (Addgene plasmid # 32726) (McCormick and Tetsu, 1999). pcDNA3‐SALL2 plasmid was described elsewhere (Escobar et al., 2015). Alexa Fluor 488‐conjugated phalloidin and Alexa Fluor 488‐conjugated goat anti‐rabbit secondary antibodies were purchased from Invitrogen (Carlsbad, CA, USA). Horseradish peroxidase‐conjugated secondary antibodies and Hoechst 33342 were from Bio‐Rad (Hercules, CA, USA).

E. Hermosilla V., Salgado G., Riffo E., Escobar D., Hepp M.I., Farkas C., Galindo M., Morín V., García‐Robles M.A., Castro A.F, & Pincheira R. (2018). SALL2 represses cyclins D1 and E1 expression and restrains G1/S cell cycle transition and cancer‐related phenotypes. Molecular Oncology, 12(7), 1026-1046.

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Whole Cell Extract Immunoblotting

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Whole cell extracts were prepared by lysing cells in two volumes of Lysis Buffer (250 mM NaCl, 5 mM ethylenediaminetetraacetic acid (EDTA), 50 mM HEPES, 0.1% v/v NP40). A total of 60 μg of whole cell extract (WCE) was resolved on 4–12% NOVEX gels using MOPS buffer (Invitrogen). Protein was transferred to PVDF membrane (Invitrogen) and blotted for BCLAF1 (BTF 608A, Bethyl Labs); THRAP3 (A300–956A, Bethyl Labs), GAPDH (ab9485, Abcam), pATM (S1981) (ab79891 Abcam), ATM (2C1, Santa Cruz), pATR (S428) (2853, Cell Signaling), ATR (N-19, Santa Cruz), 53BP1 (NB100–34, Novus Biologicals), γH2AX (S139) (JBW301, Millipore), cyclin B1 (GNS1) (sc-245, Santa Cruz), pP53 (S15) (9284S, Cell Signaling), pKAP1 (S824) (A300–767A, Bethyl Labs), FANCD2 (FI17) (sc200–22, Santa Cruz), BRCA2 (H-300) (sc-8326, Santa Cruz), FANCL (B-11) (sc-137067, Santa Cruz), PALB2 (H-45) (sc-382436, Santa Cruz) and Rad51 (3C10) (sc-53428, Santa Cruz).

Vohhodina J., Barros E.M., Savage A.L., Liberante F.G., Manti L., Bankhead P., Cosgrove N., Madden A.F., Harkin D.P, & Savage K.I. (2017). The RNA processing factors THRAP3 and BCLAF1 promote the DNA damage response through selective mRNA splicing and nuclear export. Nucleic Acids Research, 45(22), 12816-12833.

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Western Blot Analysis of Protein Signaling

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Protein lysates were lysed in Laemmli sample buffer, separated by SDS-PAGE, and proteins were transferred to polyvinylidene difluoride membranes. The following primary antibodies were used: BAP1 (#13271, RRID:AB_2798168), E-cadherin (#5296, RRID: AB_10706939), phosphop44/42 MAPK (ERK1/2) (Thr202/Tyr204) (#9101, RRID:AB_331646), S6 ribosomal protein (#2217, RRID: AB_331355), phospho-S6 ribosomal protein (Ser235/236) (#4857, RRID:AB_2181035) from Cell Signaling Technology; CADM1 (ab138697) from Abcam; b-actin (#A2066, RRID:AB_476693) from Sigma-Aldrich, phospho-Cdk1 (pTpY14/15) (#44-686G, RRID: AB_1491072) from Invitrogen; cyclin B1 (GNS-1) (#sc-245, RRID:AB_627338) and ERK1 (#sc-93, RRID:AB_631453) from Santa Cruz Biotechnology. Immunoreactivity was detected using horseradish peroxidase–conjugated secondary antibodies from (CalBioTech) and chemiluminescence substrate from ThermoFisher Scientific on a Versadoc Imaging System (Bio-Rad).

Baqai U., Purwin T.J., Bechtel N., Chua V., Han A., Hartsough E.J., Kuznetsoff J.N., Harbour J.W, & Aplin A.E. (2022). Multi-omics Profiling Shows BAP1 Loss Is Associated with Upregulated Cell Adhesion Molecules in Uveal Melanoma. Molecular cancer research : MCR, 20(8), 1260-1271.

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Antibody Production and Purification Protocol

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UCHL3 antibody was produced in collaboration with the IGBMC antibody facility using immunized rabbits and purified with SulfoLink resins according to the manufacturer's protocol. The astrin polyclonal rabbit antibody was a kind gift from Ulrike Gruneberg (Cancer Research, UK). Commercial antibodies: mouse monoclonal BubR1 (BD Biosciences, 612502 clone 9/BubR1), mouse monoclonal α-tubulin (Sigma Aldrich, T5168), human polyclonal CREST (Antibodies Incorporated, 15 234), rabbit polyclonal Aurora B (Abcam ab2254), mouse monoclonal UCHL3 (Sigma Aldrich, clone H7171), mouse monoclonal CENP-E (Thermo Scientific, MA1-5758), rabbit polyclonal GFP (Abcam, ab290)), cyclin B1 (GNS1) (Santa-Cruz, sc-245), β-actin (Sigma Aldrich, A2228), PP1γ (Santa Cruz, sc-517354), HOIP (Abcam, ab125189). MAD2L1 (GeneTex, GTX104680, survivin (Abcam, ab469).

Jerabkova K., Liao Y., Liao Y., Kleiss C., Fournane S., Durik M., Agote-Arán A., Brino L., Sedlacek R, & Sumara I. (2020). Deubiquitylase UCHL3 regulates bi-orientation and segregation of chromosomes during mitosis. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(9).

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Protein Expression Analysis by SDS-PAGE and Western Blotting

Cited 2 times

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SDS-PAGE and western blotting were performed as previously described [18 (link), 45 (link)]. The protein content for samples was determined using the BCA Protein Assay Kit (Pierce). Samples containing approximately 20–40 μg of protein were boiled in LDS sample buffer, separated on SDS-polyacrylamide gels, electrophoretically transferred to Immunoblot-P transfer membranes (IPV H00010, EMD Millipore corporation), and probed with the following primary antibodies; IRE-1 (#3294S), p-AKt (Ser 473) (#9271), AKt (#9272), pAKt (Thr 308) (#9275S), GPX (#3206S) from Cell Signaling Technology (Danvers, MA), Cyclin B1 (GNS1, #sc-245), cyclin D1 (#sc-8396), cyclin A (#H-432), PRx (A-6) P (Sc-137150P) from Santa Cruz Biotechnology (Dallas, TX), and GAPDH (#G8795) from Sigma Aldrich. The proteins were visualized using horseradish-peroxidase-conjugated secondary antibodies (#7074, Cell Signaling Technology) followed by Supersignal West Pico Chemiluminescent Substrate (#34087, Thermo Scientific). Results were quantified by densitometry of digitized images using ImageJ software (NIH, Bethesda, MD, ver.1.43) and expressed as a ratio to a GAPDH loading control.

Banerjee A., Banerjee V., Czinn S, & Blanchard T. (2017). Increased reactive oxygen species levels cause ER stress and cytotoxicity in andrographolide treated colon cancer cells. Oncotarget, 8(16), 26142-26153.

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