Cyclin a c 19

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Cyclin A (C-19) is an antibody produced by Santa Cruz Biotechnology for use in research applications. It is designed to detect the Cyclin A protein, which is a key regulator of the cell cycle and plays a critical role in cell division and proliferation.

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Lab products found in correlation

Nocodazole, by Merck Group (1 mentions) Alexa fluor 488 conjugated phalloidin, by Thermo Fisher Scientific (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) Phosphatase inhibitor cocktail 2, by Merck Group (1 mentions) Hoechst 33342, by Bio-Rad (1 mentions) Protease inhibitor cocktail 1, by Merck Group (1 mentions) Cyclin b1 gns1, by Santa Cruz Biotechnology (1 mentions) β actin ac 15, by Santa Cruz Biotechnology (1 mentions) 5 bromo 2 deoxyuridine, by Merck Group (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (1 mentions) Propidium iodide, by Merck Group (1 mentions) Rad51 h 92, by Santa Cruz Biotechnology (1 mentions) Cdk2 m2, by Santa Cruz Biotechnology (1 mentions) Nontarget shrna, by Merck Group (1 mentions) Anti cyclin d1 h 295, by Santa Cruz Biotechnology (1 mentions) Gst z 5, by Santa Cruz Biotechnology (1 mentions) Control sirna, by Qiagen (1 mentions) Cyclin d1 dcs 6, by Cell Signaling Technology (1 mentions) C myc, by Merck Group (1 mentions)

4 protocols using cyclin a c 19

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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Cyclin D1 Knockdown Protocols

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Cyclin D1-specific siRNA A (siD1-A, 5’-CCAAUAGGUGUAGGAAAUAGCGCTG-3’) was from Integrated DNA Technologies. Cyclin D1-specific siRNA B (siD1-B, 5’-AACACCAGCTCCTGTGCTGCG-3’), C (siD1-C, 5’-GCCCTCGGTGTCCTACTTCAA-3’), control siRNA (AllStars Negative control) were from Qiagen (Valencia, CA, USA). Cyclin D1 shRNA (5’-GCCAGGATGATAAGTTCCTTT-3’), and non-target shRNA (5’-CAACAAGATGAAGAGCACCAA-3’) were from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA). The following antibodies were used: anti-cyclin D1 H295, RAD51 H-92, cyclin A C-19, GST Z-5, CDK2 M-2, CDK4 C-22 antibodies (Santa Cruz Biotechnologies, Santa Cruz, CA, USA), cyclin D3 DCS-22, antibody against the C-terminus of cyclin D1 (Ab3, Thermo Fisher Scientific, Waltham, MA, USA), anti-BRCA2 OP-95 antibody (EMD Millipore), anti-HA 12CA5, CDC25A, γH2AX antibodies (abcam, Cambridge, MA, USA), anti-β actin AKR-002, GAPDH AKR-001antibodies (Sigma-Aldrich). Anti-phospho-Ser3291 BRCA2 antibody was described previously (23 (link)).

Chalermrujinanant C., Michowski W., Sittithumcharee G., Esashi F, & Jirawatnotai S. (2015). Cyclin D1 promotes BRCA2-Rad51 interaction by restricting cyclin A/B-dependent BRCA2 phosphorylation. Oncogene, 35(22), 2815-2823.

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Protein Extraction and Immunoblotting Assay

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Cells were lysed in RIPA buffer [50 mM Tris-HCl (pH 8), 150 mM NaCl, 1% (v/v) NP40, 0.5% (v/v) Na-deoxycholate, 0.1% (v/v) SDS] with 1 tablet/10 ml Complete Mini EDTA-free protease inhibitors (Roche). SDS-PAGE and immunoblots were performed using standard protocols.
Antibodies against the indicated proteins were used as follows – HIF2α (PA1-16510, Thermo Scientific), Cezanne (custom antibody, Eurogentec), β-actin (3700, Cell Signaling), PHD3 (A300-327A, Bethyl Labs), BNIP3 (ab10433, Abcam), Glut-1 (53519, Anaspec), p52 (05-361, Merck Millipore), E2F1 (3742, Cell Signaling), SP1 (07-645, Upstate-Millipore), cyclin D1 (DCS6, Cell Signaling), cyclin E (HE12, Cell Signaling), cyclin A (C-19, Santa Cruz), GFP (2956, Cell Signaling), cleaved PARP (D214) (9541, Cell Signaling), c-Myc (9E10, Sigma), phosphorylated Chk1 at S345 (2341, Cell Signaling).

Moniz S., Bandarra D., Biddlestone J., Campbell K.J., Komander D., Bremm A, & Rocha S. (2015). Cezanne regulates E2F1-dependent HIF2α expression. Journal of Cell Science, 128(16), 3082-3093.

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Antibody Characterization for Stem Cell Markers

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The antibodies used were: policlonal VRK1 (VC) and monoclonal VRK1 (1B5 or 1F6)64 (link). VRK1 (HPA000660, Sigma). Sox2 (D6D9, Cell Signaling; Berverly, MA,; E4, Santa Cruz; Santa Cruz, CA; Y-17, Santa Cruz,). Oct4 (2840, Cell Signaling), Nanog (D73G4, Cell Signaling), N-Cadherin (H-63, Santa Cruz), Vimentin (RV202, Abcam; Cambridge, UK). Flag epitope (Sigma-Aldrich, monoclonal M5; Sigma-Aldrich, polyclonal F7425). HA epitope (Santa Cruz, F-7; Sigma-Aldrich, H6908), myc epitope (06–549, Millipore; Billerica, MA; 05–724, Millipore), GST (B-14, Santa Cruz). Cyclin D1 (M-20, Santa Cruz). Cyclin A (C-19, Santa Cruz). Rb (C-15, Santa Cruz). Phospho-Rb (Ser807/811)(9308, Cell Signaling). PCNA (PC10, Santa Cruz). PAX6 (PRB-278P, Covance; Princeton, NJ), p27 (610241, BD-Transduction Laboratories; Franklin Lakes, NJ). CREB (9104, Cell Signaling). Phospho-CREB (Ser133) (9191, Cell Signaling). C-myc (N-262, Santa Cruz). β-actin (AC-15, Sigma-Aldrich). Secondary antibodies goat α-Mouse IgG, DyLightTM 680 and/or goat α-Rabbit Ig-G, DyLightTM 800 (Thermo Fischer Scientific) were used for detection in a Li-Cor Odyssey system (Thermo Fisher Scientific; Waltham, MA).

Moura D.S., Fernández I.F., Marín-Royo G., López-Sánchez I., Martín-Doncel E., Vega F.M, & Lazo P.A. (2016). Oncogenic Sox2 regulates and cooperates with VRK1 in cell cycle progression and differentiation. Scientific Reports, 6, 28532.

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