Anti-YAP (Cat# 14074S, RRID: AB_2650491 ), anti-Phospho-YAP (Ser127) (Cat# 13008S, RRID: AB_2650553 ), anti-Cyclin D1 (Cat# 2922S, RRID: AB_2228523 ), anti-Cyclin D1 (Cat# 55506S, RRID: AB_2827374 ), anti-Cyclin E1 (Cat# 20808, RRID: AB_2783554 ), anti-β-actin (Cat# 4970S, RRID, AB_2223172), anti-Lamin B1 (Cat# 13435S, RRID: AB_2737428 ), anti-GAPDH (Cat# 2118S, RRID: AB_561053 ), anti-rabbit, IgG HRP-linked antibody (Cat# 7074S, RRID: AB_2099233 ), anti-mouse IgG, HRP-linked antibody (Cat# 7076S, RRID: AB_330924), signalstain® boost IHC detection reagent (HRP, rabbit) (Cat# 8114S, RRID: AB_10544930), signalstain® boost IHC detection reagent (HRP, mouse) (Cat# 8125S, RRID: AB_10547893) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-CCNA1 (Cat# D220507, RRID: AB_2819214 ), anti-CYR61 (Cat# D122190, RRID: AB_2819221 ), anti-ANKRD1 (Cat# D121628, RRID: AB_2819216 ) and anti-CTGF (Cat# D160212, RRID: AB_2819217 ) were purchased from Sangon Biotech (Shanghai, China). Anti-β-catenin (Cat# 610153, RRID: AB_397554 ) was purchased from BD Biosciences (Franklin Lakes, NJ, USA). Anti-caspase-3 (Cat# sc-56053, RRID: AB_781826 ) was purchased from Santa Cruz Biotechnology, Inc. (Carpinteria, CA, USA).
Anti cyclin d1
Manufactured by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Canada
Anti-cyclin D1 is a laboratory reagent used to detect the presence and expression levels of the cyclin D1 protein. Cyclin D1 is a key regulator of the cell cycle and plays a crucial role in the progression of cells through the G1 phase. The anti-cyclin D1 reagent can be used in various experimental techniques, such as western blotting, immunohistochemistry, and flow cytometry, to study the expression and regulation of cyclin D1 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Cell Signaling Technology (48 mentions)
Anti cdk4, by Cell Signaling Technology (46 mentions)
Anti bcl 2, by Cell Signaling Technology (45 mentions)
Anti gapdh, by Cell Signaling Technology (41 mentions)
Anti akt, by Cell Signaling Technology (39 mentions)
Anti p21, by Cell Signaling Technology (39 mentions)
Anti β catenin, by Cell Signaling Technology (35 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (33 mentions)
Anti c myc, by Cell Signaling Technology (32 mentions)
Anti vimentin, by Cell Signaling Technology (29 mentions)
Anti bax, by Cell Signaling Technology (27 mentions)
Anti parp, by Cell Signaling Technology (26 mentions)
Anti cdk6, by Cell Signaling Technology (25 mentions)
Anti e cadherin, by Cell Signaling Technology (22 mentions)
Anti cyclin b1, by Cell Signaling Technology (21 mentions)
Anti p27, by Cell Signaling Technology (21 mentions)
Anti cdk2, by Cell Signaling Technology (20 mentions)
Anti p akt, by Cell Signaling Technology (19 mentions)
Anti caspase 3, by Cell Signaling Technology (18 mentions)
Anti gapdh, by Santa Cruz Biotechnology (16 mentions)
332 protocols using anti cyclin d1
1
Signaling Pathway Antibody Panel
2
CD58 and Akt Modulators in Stem Cell
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Soluble CD58 (sCD58), AKT inhibitor (LY294002), and Akt activator (SC-79) were acquired from Med Chem Express (MCE, Shanghai, China). Anti-Oct4 (11263-1-AP), anti-Sox2 (66411-1-Ig), anti-CD24 (18330-1-AP), anti-vimentin (10366-1-AP), and anti-c-Myc (10828-1-AP) were purchased from Proteintech (Wu Han, China). Anti-EPCAM (#2626S), anti-E-cadherin (#3195S), anti-AKT (#9272S), anti-GSK-3β (#5676S), anti-Phospho-GSK-3β (Ser9) (5558S), anti-β-Catenin (#8480S), anti-non-phosphorylated (active) β-catenin (S33/37/T41) (#8814S), anti-phospho-β-Catenin (Ser552) (#9566S), and anti-cyclin D1 (#55506S) were acquired from Cell Signaling Technology. Anti-CD58 (A0806) and anti-phospho-AKT (Ser473) (AP0140) were purchases from ABclonal (Wu Han, China).
3
Western Blot Analysis of Cell Signaling Proteins
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Protein sample preparation and western blot assay were performed as previously described (24 (link)). The primary antibodies used were: anti-MTMR3 (1:500; cat. no. 12443; Cell Signaling Technology, Inc.), anti-Cyclin D1 (1:1,000; cat. no. 60186-1-1g; ProteinTech Group, Inc.), anti-cyclin-dependent kinase 2 (1:1,000; CDK2; cat. no. 11026-2-AP; ProteinTech Group, Inc.), anti-p62 (1:1,000; cat. no. 19117-1-AP; ProteinTech Group, Inc.), anti-p21 (1:1,000; cat. no. 2947; Cell Signaling Technology, Inc.), anti-Cyclin E (1:1,000; cat. no. sc-247; Santa Cruz Biotechnology, Inc.), anti-Cyclin A (1:1,000; cat. no. sc-751; Santa Cruz Biotechnology, Inc.), anti-cell division cycle 25 A (1:1,000; cdc25A; sc-7157; Santa Cruz Biotechnology, Inc.), anti-microtubule associated protein 1 light chain 3 (LC3) A (1:500; cat. no. 4599; Cell Signaling Technology, Inc.), anti-LC3B (1:500; cat. no. 3868; Cell Signaling Technology, Inc.) and anti-GAPDH (1:20,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.). Bound HRP-labeled secondary antibody (1:5,000; cat. no. SC-2005 or SC-2054; Santa Cruz Biotechnology, Inc.) was assayed by super ECL detection reagent (Pierce; Thermo Fisher Scientific, Inc.). Protein density of western blots was analyzed using ImageJ 1.51k software (National Institutes of Health).
4
Western Blot Analysis of Protein Markers
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Western blots were performed as previously described [11] (link). Briefly, as soon as the harvested cells were lysed and the supernatant was collected, 60 µg protein was loaded onto SDS-PAGE and transferred to polyvinylidene fluoride membranes (PVDF). Membranes were blocked with 5% skimmed milk for 1 hour and incubated overnight with one of the following primary antibodies: anti-PSMD10 (diluted at 1∶1,000; Sigma, St. Louis, MO, USA), anti-Twist2 (diluted at 1∶200; Abcam, Cambridge, UK), anti-Vimentin (diluted at 1∶500; Cell Signaling Technology, Beverley, MA, USA), anti-β-catenin (diluted at 1∶500; Cell Signaling Technology), anti-E-cadherin (diluted at 1∶500; Cell Signaling Technology) and anti-cyclin D1(diluted at 1∶500; Cell Signaling Technology) rabbit monoclonal antibody, followed by 1 hour of incubation with the appropriate secondary antibody (1∶5,000). The anti-GAPDH (Epitomics) rabbit monoclonal antibody was diluted to 1∶1,000 for use as a sample loading control.
5
Cell Lines and Reagents for Cancer Research
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HEK 293T, HCT116, HT29, LoVo, SW480, SW620 and RKO cells were obtained from our collaborators and maintained in DMEM (GIBCO) supplemented with 10% FBS (HyClone). CSBF/C10orf99 was synthesized in Chinese Peptide Company (Hangzhou, China) with a purity > 97%. Gal1 was purchased from R&D Systems (1152-GA-050). Antibodies used in this paper were: mouse anti-myc (9E10) (# SAB4700447, Sigma-Aldrich), rabbit anti-HA (C29F4) (# 3724, Cell Signaling Technology), goat anti-Human IgG (Fc specific) (# I2136, Sigma-Aldrich), anti-cyclin D1 (# 2926, Cell Signaling Technology), anti-cyclin D3 (# 2936, Cell Signaling Technology), anti-CDK4 (# 2906, Cell Signaling Technology), anti-CDK6 (# 3136, Cell Signaling Technology), anti-phospho-Cyclin B1 (Ser147) (# 4131, Cell Signaling Technology), anti-SUSD2 rabbit polyclonal antibody (# HPA004117, Sigma-Aldrich), anti-β-tublin (# T8328, Sigma-Aldrich) and anti-GAPDH (# 2118, Cell Signaling Technology). Primary cancer tissues and paired adjacent tissues were obtained from patients under primary surgery at Peking University Cancer Hospital (Beijing, China), with patients' consent and institutional ethics approval. Fresh human tissues were fixed with 10% formalin in PBS for immunohistochemistry, or frozen in liquid nitrogen for RNA extraction. This investigation was carried out after approval by the Ethics Committee of Peking University Cancer Hospital.
6
Protein Expression Analysis in GMCs
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GMCs were lysed in RIPA lysis buffer (Beyotime, Shanghai, China). Total proteins were separated by sodium dodecyl sulfate/polyacrylamide gel electrophoresis, transferred to polyvinylidenefluoride membrane, and blocked with 5% skim milk for 2 h. Then the membrane was incubated with specific primary antibodies (rabbit anti-mouse; anti-PSMD11, #14303; anti-proliferating cell nuclear antigen (PCNA), #13110; anti-inhibitor of NF-κB α (IκBα), #4812; anti- phosphorylated IκBα (p-IκBα), #2859; anti-NF-κB p65, #8242; anti-Histone H3, #9728; anti-COX-2, #4842; anti-cyclinD1, #2922; anti-GAPDH, #5174; 1:1000, Cell Signaling Technology, Boston, U.S.A.) for 12 h at 4°C. After incubating with horseradish peroxidase (HRP)–conjugated secondary antibody (goat anti-rabbit; 1:2000, Cell Signaling Technology) for 1 h at 25°C, the protein brands were visualized using an HRP color development kit (Thermo Fisher Scientific). The protein level was normalized to GAPDH, and standardized to L-GMC group (set as 1).
7
Western Blot Analysis of Protein Expression
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Anti-FLAG and anti-beta actin (Sigma-Aldrich, USA); anti-GNL1, anti-RPS20, anti-CDK1 and anti-MDM2 (Abcam, UK); anti-GFP, anti-Cyclin B1 and anti-p53 (Santa Cruz, USA); anti-p21, anti-Cyclin D1, anti-CDK4, anti-pRbSer-780, anti-Rb and anti-E2F1 (Cell Signaling Technology Inc., USA) antibodies were used in western blot analysis for checking protein expression.
8
Baicalin and 3-MA Modulate Autophagy and Apoptosis
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Baicalin and 3-MA were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). BAPTA-AM was purchased from Selleck Chemicals (Shanghai, China). Anti-Bax, anti-Bcl-xl, anti-cleaved caspase 3, anti-PARP, anti-mTOR, anti-p-mTOR, anti-AKT, anti-p-AKT, anti-cyclin D1, anti-cyclin B1, anti-cyclin A, anti-ZO-1, anti-Catenin, anti-Vimentin, and anti-β-actin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-LC3B, anti-MMP-2, and anti-beclin 1 antibodies were purchased from Novus Biologicals (Littleton, CO, USA). Anti-HMGB1 and anti-p62/SQSTM1 antibodies were purchased from Abcam (Cambridge, MA, USA). Horseradish peroxidase (HRP)-conjugated and FITC-conjugated antirabbit or antimouse IgG antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).
9
Overexpression and Silencing of PRSS8
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The full length of PRSS8 was cloned from human cDNA and inserted into pEGFP vector (Promega, Madison, WI), to generate pEGFP -PRSS8, expression constructs. A 19-nt siRNA oligonucleotide with 3′-dt extensions against human PRSS8 transcript and one scrambled siRNA (negative control) were designed, as shown in Supplementary Table S1 . siRNAs were synthesized by Shanghai GenePharma Inc. (Shanghai, China). Twenty-four hours before transfection, 1.0×105 cells were seeded in a 6-well plate. 4 μg of PRSS8 expression plasmid or negative control plasmid was transfected into cells, using Lipofectamine 3000 (Invitrogen, Carlsbad, CA) following the manufacture's protocol. The cells were collected for immunoblotting analysis using the following primary antibodies: anti-PRSS8, anti-P21, anti-Cyclin D1, anti-E-cadherin, anti-snail and anti-Twist (from Cell Signaling Technologies Inc., CA). Anti-β-actin (Sigma, St Louise, MO) was used as internal loading control.
10
Western Blot Protein Detection
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Total proteins were extracted using 1X cell lysing buffer (Cell Signaling Technology). Samples were separated by electrophoresis in SDS- polyacrylamide gels (12%) and transferred to PVDF membranes. Membranes were blocked for 1 h with PBST (PBS plus 0.1% Tween-20) containing 5% non-fat milk. Blots were incubated overnight at 4°C with primary antibodies in PBST containing 5% BSA at the manufacturer's recommended dilution. The antibodies were purchased from the following suppliers: FKBP4 (ProteinTech); anti-pS473 Akt, anti-pT308 Akt, anti-Akt, anti-pT286 Cyclin D1, anti-Cyclin D1, anti-E2F-1, anti-pT37/46 4E-BP1, anti-pS9 GSK-3β, anti-GSK-3β, and anti-mTOR (Cell Signaling Technology); anti-GAPDH and anti-Actin (Santa Cruz Biotechnology); anti-Tubulin and anti-PIK3R2 (Sigma-Aldrich). After washing, blots were incubated with either an anti-rabbit HRP, or an anti-mouse HRP-conjugated antibody (Cell Signaling Technology) for 1 h at 25°C. After washing, blots were developed with Supersignal WEST Pico Plus (Life Technologies SAS).
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