Cyclin d1

Manufactured by Cell Signaling Technology

Sourced in United States, China, United Kingdom, Germany, Japan

Cyclin D1 is a key regulatory protein involved in cell cycle progression. It plays a crucial role in the G1/S transition phase of the cell cycle. Cyclin D1 functions as a regulatory subunit of cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6), promoting cell cycle advancement.

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Lab products found in correlation

Close spelling variants of this product

Cyclin D1 (E3P5S) Cyclin D1 (E3P5S) XP® Rabbit mAb Antibody for cyclin D1 Antibodies against cyclin D1 Rabbit anti-Cyclin D1 monoclonal antibody Rabbit antibodies against Cyclin D1 Cyclin D1(2922) Cyclin D1 (55506), Cyclin D1 (CCND1) Phospho-cyclin D1

991 protocols using cyclin d1

Silibinin and NAC Mechanism Study

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Silibinin (purity >98%, as assessed by high-performance liquid chromatography) and N-acetyl-L-cysteine (NAC) were procured from Sigma-Aldrich (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO) was used to dissolve Silibinin during in vitro experiments. Primary antibodies against Bcl-2 (#3498), cleaved PARP (#5625), Bax (#5023), cyclin D1 (#2922), cleaved caspase-3 (#9661), cyclin-dependent kinase4 (CDK4) (#12790), CDK6 (#3136), cyclin E1 (#4129), E-cadherin (#3195), N-cadherin (#4061), , p-AKT (#9271), AKT (#9272), p-ERK (#9101), ERK (#9102), p-p38 (#9216), p38 (#9212), cyclin D1 (#2922), p-c-Jun (#3270S), and c-Jun (#9165) were procured from Cell Signaling Technology (Danvers, MA, USA). And p53 (sc-1312), SOD1 (sc-11407), SOD2 (sc-18503), β-Actin (sc-47778) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Also, vimentin (ab92547) was obtained from abcam.

Zhang H., Kim H., Kim S.Y., Hai H., Kim E., Ma L., Kim D., Kim C.Y., Park K., Park S., Ko J., Kim E.K., Kim K., Ryoo Z.Y., Yi J, & Kim M.O. (2023). Silibinin induces oral cancer cell apoptosis and reactive oxygen species generation by activating the JNK/c-Jun pathway. Journal of Cancer, 14(10), 1875-1887.

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Nrf2 Knockdown Impact on Target Genes

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To determine the impact of knocking down Nrf2 on its target genes
HO1 and NQO1 protein expression, cell lysates were collected and analyzed
on 10% NuPAGE gels as detailed in Madhunapantula et al.^{58 (link)} Total protein quantity was determined using
the BCA assay.^{59 (link)} About 50 μg of total
protein/well was loaded and analyzed as described in Madhunapantula
et al.^{58 (link)} Expression of Nrf2, HO1, NQO1
and p53, Bax, Cyclin-D1, and P27 was measured by detecting the respective
proteins using primary antibodies recognizing Nrf2 (cat no. 12721),
NQO1 (cat no. 62262), HO1 (cat no. 5853), p53 (cat no. 9282S), Bax
(cat no. D2E11), Cyclin-D1 (cat no. SC718), and P27 (cat no. SC 393380)
obtained from Cell Signaling Technologies, Danvers, MA. Enolase (cat
no. SC-7455) and secondary antibodies (antirabbit and antigoat) conjugated
with horseradish peroxidase (HRP; rabbit cat no. SC2357 and goat cat
no. SC2020) were from Santacruz Biotechnology, Dallas, TX. The proteins
were detected using ECL (Western Bright ECL cat no. K-12045-D20),
Advansta Corporation, San Jose, CA.

Chikkegowda P., Pookunoth B.C., Bovilla V.R., Veeresh P.M., Leihang Z., Thippeswamy T., Padukudru M.A., Hathur B., Kanchugarakoppal R.S., B.a.s.a.p.p.a, & Madhunapantula S.V. (2021). Design, Synthesis, Characterization, and Crystal Structure Studies of Nrf2 Modulators for Inhibiting Cancer Cell Growth In Vitro and In Vivo. ACS Omega, 6(15), 10054-10071.

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Cellular Signaling Pathway Analysis

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SNU484 and SNU638 were acquired from the Korean Cell Line Bank (Seoul National University, Korea). RPMI-1640 medium (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) (Welgene Gold Serum, Gyeongsan-si, Republic of Korea) and 1% penicillin (Sigma-Aldrich, St. Louis, USA) were used for cell culture. Antibodies, such as glyceraldehyde 3-phosphate dehydrogenase (GAPDH), cleaved-PARP, cleaved-caspase-9, PARP, caspase 9, cyclin D1, Akt, p-Akt, GSK-3β, β-catenin, p-β-catenin, cyclin D1, and lamin B, and secondary antibodies against rabbit and mice were acquired from Cell Signaling Technology, Inc. (Danvers, Massachusetts, USA); E-cadherin, uPA, MMP-9, and c-Myc were bought from Santa Cruz Biotechnology, Inc. (Santa CRUZ, CA, USA). Wortmannin and 5-FU were acquired from Sigma-Aldrich (St. Louis, MO, USA) and DIM was acquired from LKT Laboratories (St. Paul, MN, USA).

Li C.S., Nguyen T.V., Chai O.H., Park B.H., Lee J.S, & Kim S.M. (2023). 3,3′-Diindolylmethane Augments 5-Fluorouracil-InducedGrowth Suppression in Gastric Cancer Cells through Suppression of the Akt/GSK-3β and WNT/Beta-Catenin. Journal of Oncology, 2023, 8268955.

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Wnt Signaling Pathway Modulation

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HCT-116 and SW480 cells were maintained in culture as described above. The cells were plated in 6-well plates and incubated at the same time alongside cells used for analysis of β-catenin. The media was then replaced with fresh media containing the indicated compounds in DMSO or DMSO. The final DMSO concentration was 0.1%. The cells were incubated for 18 hours. The media was removed, the cells washed with PBS, and then lysed with 240μL 1X Laemmli sample buffer. Immunoblots using antibodies to c-Myc (Santa Cruz Biotechnology, SC-789), Cyclin D1, and Survivin (Cell Signaling Technology, cat#2978 and 2808) were used to detect c-Myc, Cyclin D1 and Survivin levels in the total cell lysates. Antibodies to β-actin (C-4, Santa Cruz Biotechnology, SC-47778) was used to measure levels of β-actin, which served as a loading control in all immunoblots.

Mook RA J.r., Ren X.R., Wang J., Piao H., Barak L.S., Lyerly H.K, & Chen W. (2017). Benzimidazole inhibitors from the Niclosamide chemotype inhibit Wnt/β-catenin signaling with selectivity over effects on ATP homeostasis. Bioorganic & medicinal chemistry, 25(6), 1804-1816.

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Histochemical Analysis of Intestine

Cited 1 time

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The intestine was processed for histochemical analysis as described (Cox et al., 2018 (link)). Antibodies used were: Cyclin D1 (cat. 2978; Cell Signaling), E-cadherin (cat. 3195; Cell Signaling), Ki67 (cat 15580; Abcam), OLFM4 (cat. 39141; Invitrogen), Chromogranin A (Chgr A) (cat. ab15160; abcam), rabbit anti-HES1 (cat. 11988; CST) and anti-EDTB hybridoma supernatant (Wilson et al., 1987 (link)). Antigen retrieval with Tris/EDTA pH 9 was used for Cyclin D1 (1:50). Citrate pH 6 antigen retrieval was used for E-cadherin (1:200), Ki67 (1:200), Chgr A (1:200), HES1 (1:200) and OLFM4 (1:400). Signal Boost (cat. 8114S; Cell Signaling Technology) and DAB detection kits (cat. 8059S; Cell Signaling Technology) were used for visualization of Cyclin D1 and HES1. To visualize goblet cells, tissue was labelled with Alcian Blue Solution according to manufacturer’s instructions (cat. IW-3000A; IHCWORLD).

Wu M.H., Padilla-Rodriguez M., Blum I., Camenisch A., da Paz V.F., Ollerton M., Muller J., Momtaz S., Mitchell S.A., Kiela P., Thorne C., Wilson J.M, & Cox C.M. (2021). Proliferation in the developing intestine is regulated by the endosomal protein Endotubin. Developmental biology, 480, 50-61.

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Protein Expression Profiling by Western Blot

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Western blot assay was used to detect protein expression levels. First, cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (25 mmol/L Tris∙HCl pH7.6, 150 mmol/L NaCl, 1% NP‐40, 0.25% sodium deoxycholate, 0.1% SDS) with protease inhibitor cocktail (Roche Diagnostics, Mannheim, Germany). Then, the protein lysates were denatured at 95°C for 5 minutes after mixing with 5x SDS‐loading buffer. Subsequently, the cell extracts (30 µg protein) were separated on a sodium dodecyl sulphate‐polyacrylamide electrophoretic gel (SDS‐PAGE) and then transferred to nitrocellulose membranes. After blocked with 3% BSA for 2 hours, the membranes were incubated overnight at 4°C with the following primary antibodies at dilutions of 1:1000: PARP (#9532), Bcl‐2 (#2876), Bax (#14796), Caspase‐9 (#9502), Caspase‐3 (#9664), CyclinD1 (#2978), CDK2 (#2546), CDK4 (#12790), CDK6 (#13331), CyclinD1 (#2978) purchased from Cell Signaling Technology, and PCNA (#ab29) obtained from Abcam. Next, the membrane was incubated with the corresponding horseradish peroxidase–labelled secondary antibody (Santa Cruz Biotechnology) for 2 hours at room temperature. Lastly, the signal was visualized by an enhanced chemiluminescence (ECL) kit (Immobilon Western HRP, MILLIPORE, USA) according to the manufacturer's instructions.

Zhao T., Wang C., Huo X., He M, & Chen J. (2020). Pterostilbene enhances sorafenib’s anticancer effects on gastric adenocarcinoma. Journal of Cellular and Molecular Medicine, 24(21), 12525-12536.

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Apoptosis and Cell Cycle Regulation Antibodies

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The following antibodies were all obtained from Cell Signaling Technology; catalog numbers are in parentheses: caspase 3 (9662), caspase 8 (9746), cleaved caspase 9 (9501), apoptosis-inducing factor (4642), cyclin-dependent kinase (CDK) 1 (9112), CDK2 (2546), CDK4 (2906), cyclin B1 (4135), cyclin D1 (2926), cyclin E1 (4129), p21 (2947), p27 (3686), p53 (9282), checkpoint kinase 2 (CHEK2) (6334), ataxia telangiectasia mutated (ATM) protein kinase (2873), phospho-histone H3 (pH3; Ser10) (9701), and phospho-histone H2A variant X (γ-H2AX) (9718). Horseradish peroxidase-conjugated secondary antibodies for western blot were from Santa Cruz Biotechnology (sc-2357). Alexafluor 488-conjugated secondary antibodies for immunofluorescence were from Jackson ImmunoResearch (211-545-109). Protease inhibitor cocktail (539131) and phosphatase inhibitor cocktail (524625) were both from Calbiochem. Annexin V (A13201) was from Thermo Fisher. Etoposide (GR-307) was from BioMol. Muse^® cell analyzer kits for measurement of caspase3/7 (MCH100108) and cell cycle (MCH100106) were from EMD Millipore. Unless otherwise noted, protein was measured by the method of Lowry et al. (9 (link)). Statistical significance was determined using Student’s t-test.

Yang H., Shi X., Kolar E.A., Clay E.M., Xia S., Pei Z, & Watkins P.A. (2023). VERY LONG-CHAIN ACYL-CoA SYNTHETASE-3 (ACSVL3) PROMOTES THE MALIGNANT GROWTH BEHAVIOR OF U87 GLIOMA CELLS VIA CHANGES IN CELL CYCLE WITHOUT AFFECTING APOPTOSIS. bioRxiv.

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Western Blot Analysis of Cell Signaling

Cited 2 times

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Western blotting was performed using a SDS-PAGE Electrophoresis System according to the previous description [16 (link), 17 (link)] with antibodies specific for C-myc (Cell Signaling Technology, Beverly, MA, USA), CDK4 (Cell Signaling Technology), CDK6(Cell Signaling Technology), CyclinD1(Cell Signaling Technology), Rb (Cell Signaling Technology), p-Rb (Cell Signaling Technology), Caspase3 (Immunoway, USA), Cleaved Caspase3 (Cell Signaling Technology), Snail (Proteintech, USA), Slug (Proteintech), E-cadherin (Cell Signaling Technology), N-cadherin (Cell Signaling Technology), PI3K (Abclonal Technology), p-PI3K(Cell Signaling Technology), AKT(Cell Signaling Technology), p-AKT(Cell Signaling Technology), PTTG1(Cell Signaling Technology), β-Tublin (Cell Signaling Technology) and β-actin (Proteintech).Signals were detected using enhanced chemiluminescence reagents (Millipore, Schwalbach/Ts., Germany).

Huang J.L., Cao S.W., Ou Q.S., Yang B., Zheng S.H., Tang J., Chen J., Hu Y.W., Zheng L, & Wang Q. (2018). The long non-coding RNA PTTG3P promotes cell growth and metastasis via up-regulating PTTG1 and activating PI3K/AKT signaling in hepatocellular carcinoma. Molecular Cancer, 17, 93.

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Bazedoxifene Modulates Colon Cancer Signaling

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Human colon cancer cell lines (DLD-1, HCT-15, and HCT-116) at 50–60% confluence were harvested after an overnight treatment with bazedoxifene or DMSO, and then lysed in cold RIPA lysis buffer containing protease inhibitor cocktail and phosphatase inhibitor cocktail. The lysates were subjected to 10% or 12% SDS-PAGE gel and transferred to a PVDF membrane. Membranes were probed with a 1:1000 dilution of specific primary antibody and 1:10,000 HRP-conjugated secondary antibody. Primary antibodies against phosphorylated STAT3 (Tyr705, p-STAT3^Y705), IL-6, BCL-XL, c-MYC, survivin, cyclin D1, STAT3, AKT, ERK, phospho-specific extracellular signal-regulated kinase (ERK) 1/2 (threonine 202/Tyrosine 204), phosphorylated-AKT (Ser473), GAPDH and secondary antibodies were all from Cell Signaling Technology (Beverly, MA, USA). Primary antibodies against IL-11, IL-11Rα and IL-6R were from Abcam (Cambridge, MA, USA). Membranes were analyzed using enhanced chemiluminescence plus reagents and scanned with the Storm Scanner (Amersham Pharmacia Biotech Inc., Piscataway, NJ).

Wei J., Ma L., Lai Y.H., Zhang R., Li H., Li C, & Lin J. (2019). Bazedoxifene as a novel GP130 inhibitor for Colon Cancer therapy. Journal of Experimental & Clinical Cancer Research : CR, 38, 63.

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Cell Proliferation and Apoptosis Assay

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A cell counting kit (CCK) for WST assay was purchased from Donginbiotech Co. (Seoul, Korea). Crystal violet solution was purchased from Sigma-Aldrich (St Louis, MO, USA). Anti-phospho-AMPK, phospho-p38, phospho-ERK, phospho-JNK, cyclin D1, CDK4, AMPK, PARP, caspase-3, Bcl-2, Bcl-xL, and Bax antibodies were purchased from Cell Signaling (Danvers, MA, USA). Anti-p38, ERK, JNK, GAPDH, and α-tubulin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Mun J.G., Jeon H.D., Yoon D.H., Lee Y.S., Park S.Y., Jin J.S., Park N.J, & Kee J.Y. (2022). Supercritical Extract of Cannabis sativa Inhibits Lung Metastasis in Colorectal Cancer Cells by Increasing AMPK and MAPKs-Mediated Apoptosis and Cell Cycle Arrest. Nutrients, 14(21), 4548.

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