The EdU kit (Cat No: C10310-1) was purchased from RiboBio, Guangzhou, China, and the extreme ultrasensitive ECL chemiluminescence kit (Cat No: P0018FM) was purchased from Beyotime, China. Trypsin (Cat No: T1300), MTT reagent (Cat No: M8180), cell-grade dimethyl sulfoxide DMSO (Cat No: D8371) and Rainbow 180 Broad Spectrum Protein Marker (Cat No: PR1910) were purchased from Solarbio, Beijing, China. DMEM high glucose medium (Cat No: SH30243.01) was purchased from Gibco, New York, NY, USA, and 100 U penicillin–streptomycin solution (Cat No: SV30010) was purchased from Hyclone, Los Angeles, CA, USA. PVDF membranes, 5-fluorouracil (5-FU) and Entinostat (Ent) were obtained from MERCK, Darmstadt, Germany. Bcl2 antibody (Cat No: 68103-1-Ig), Caspase-3 antibody (Cat No: 66470-2-Ig), Cytochrome C (Cyt C) antibody (Cat No: 10993-1-AP), p53 antibody (Cat No: 60283-2-Ig), p21 antibody (Cat No: 10355-1-AP), CDK2 antibody (Cat No: 10122-1-AP), HDAC1 antibody (Cat No: 66085-1-Ig), HDAC2 antibody (Cat No: 67165-1-Ig), HDAC3 antibody (Cat No: 67151-1-Ig) and HDAC8 antibody (Cat No: 17548-1 AP) were purchased from Proteintech, Wuhan, China. H4 antibody (Cat No: PTM-1009), H3K18 antibody (Cat No: PTM-114), H3K27 antibody (Cat No: PTM-5010) and H3K9 antibody (Cat No: PTM-7094) were purchased from Jingjie PTM BioLab, Hangzhou, China.
P53 antibody
Manufactured by Proteintech
Sourced in China
The P53 antibody is a laboratory reagent used for the detection and analysis of the p53 protein in various biological samples. The p53 protein is a tumor suppressor that plays a critical role in regulating cell growth and division. The P53 antibody can be used in techniques such as Western blotting, immunohistochemistry, and immunoprecipitation to identify and quantify the p53 protein.
Automatically generated - may contain errors
Lab products found in correlation
Bcl 2 antibody, by Proteintech (3 mentions)
Bax antibody, by Proteintech (3 mentions)
P21 antibody, by Proteintech (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Caspase 3 antibody, by Proteintech (1 mentions)
Hdac2 antibody, by Proteintech (1 mentions)
Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Solarbio (1 mentions)
Mtt reagent, by Solarbio (1 mentions)
5 fluorouracil 5 fu, by Merck Group (1 mentions)
Ki67 antibody, by Abcam (1 mentions)
Anti caspase 3 antibody, by Cell Signaling Technology (1 mentions)
E cadherin antibody, by Proteintech (1 mentions)
Vimentin antibody, by Proteintech (1 mentions)
Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Pcna antibody, by Proteintech (1 mentions)
Mmp 9 antibody, by Proteintech (1 mentions)
Akt antibody, by Proteintech (1 mentions)
6 protocols using p53 antibody
1
Detailed Biochemical Protocol for Cell Line Experiments
2
Rapamycin and 5-FU Synergistic Effects on Colorectal Cancer
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Rapamycin and 5-FU were obtained from meilunbio (Dalian, China). The anti Caspase3 antibody, Cleaved Caspase3 antibody, p-AKT (ser 473) antibody, p-AKT (Thr 308) antibody, p70 S6 Kinase antibody, and p-p70S6 Kinase (Thr 389) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). The ki-67 antibody was purchased from Abcam (Cambridge, UK). The BCL-2 antibody, Bax antibody, AKT antibody, p53 antibody, E-Cadherin antibody, N-Cadherin antibody, Vimentin antibody, MMP-9 antibody and PCNA antibody were purchased from Proteintech (Wuhan, China). The GAPDH rabbit polyclonal antibody was obtained from XianZhi Biotech (Hangzhou, China). The peroxidase-conjugated secondary antibodies were obtained from CWBIO (Taizhou, China).
The human CRC cell line HCT-116 and SW-480 cells were purchased from iCell Bioscience Inc (Shanghai, China). The cells were cultured in a DMEM medium containing 10% FBS in a 37 °C incubator containing 5% CO2.
Female athymic nude mice (6–8 weeks old) were purchased from Guangdong Medical Laboratory Animal Center. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals. The Laboratory Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University approved all experimental protocols.
The human CRC cell line HCT-116 and SW-480 cells were purchased from iCell Bioscience Inc (Shanghai, China). The cells were cultured in a DMEM medium containing 10% FBS in a 37 °C incubator containing 5% CO2.
Female athymic nude mice (6–8 weeks old) were purchased from Guangdong Medical Laboratory Animal Center. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals. The Laboratory Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University approved all experimental protocols.
3
Apoptosis Protein Expression Analysis
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Tissues and cells were lysed with RIPA lysis buffer containing 1 mM PMSF (Solarbio) and
protein concentration was quantified using a BCA kit (Solarbio). Proteins were
fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and
transferred to PVDF membranes (Millipore, Billerica, MA, USA). PVDF membranes were then
blocked with 5% skim milk (Sangon Biotech, Shanghai, China) and incubated with one of the
primary antibodies overnight at 4°C. After three-times PBS washing, PVDF membranes were
incubated with certain secondary antibody for 60 min at 37°C. Primary antibodies used in
this study were as follows: p53 antibody (1:2,000, Proteintech, Hangzhou, China), p21
antibody (1:1,000, Abcam, Cambridge, UK), cleaved caspase 3 antibody (1:1,000, CST,
Framingham, MA, USA), cleaved caspase 9 antibody (1:1,000, CST), BCL-2 antibody (1:2,000,
Proteintech), BAX antibody (1:5,000, Proteintech), and GAPDH antibody (1:10,000,
Proteintech). HRP-conjugated goat anti-rabbit lgG (1:3,000, Solarbio) and HRP-conjugated
goat anti-mouse lgG (1:3,000, Solarbio) were used as secondary antibodies.
protein concentration was quantified using a BCA kit (Solarbio). Proteins were
fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and
transferred to PVDF membranes (Millipore, Billerica, MA, USA). PVDF membranes were then
blocked with 5% skim milk (Sangon Biotech, Shanghai, China) and incubated with one of the
primary antibodies overnight at 4°C. After three-times PBS washing, PVDF membranes were
incubated with certain secondary antibody for 60 min at 37°C. Primary antibodies used in
this study were as follows: p53 antibody (1:2,000, Proteintech, Hangzhou, China), p21
antibody (1:1,000, Abcam, Cambridge, UK), cleaved caspase 3 antibody (1:1,000, CST,
Framingham, MA, USA), cleaved caspase 9 antibody (1:1,000, CST), BCL-2 antibody (1:2,000,
Proteintech), BAX antibody (1:5,000, Proteintech), and GAPDH antibody (1:10,000,
Proteintech). HRP-conjugated goat anti-rabbit lgG (1:3,000, Solarbio) and HRP-conjugated
goat anti-mouse lgG (1:3,000, Solarbio) were used as secondary antibodies.
4
Protein Extraction and Western Blot Analysis of HCC Cells
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Total protein was extracted from HCC cells using RIPA lysis buffer and quantified using Pierce BCA Protein Assay kit according to the manufacturer's protocol (Thermo Fisher Scientific, Waltham, Massachusetts, USA). Protein extracts (10 μg) were separated by SDS/PAGE gel (Thermo Fisher Scientific) and then transferred to a nitrocellulose membrane (Thermo Fisher Scientific). After membrane was blocked with 5% non-fat dry milk dissolved in Tris-buffered saline with Tween-20 buffer for 1 h at room temperature, it was incubated at 4°C overnight with primary antibodies against RFX5 (1:1000; Proteintech), YWHAQ (1:10,000; Proteintech), p53 antibody (1:2000; Proteintech), and Bax antibody (1:1500; Proteintech) followed by incubation with HRP-conjugated secondary antibodies (1:5000; Proteintech) for 1 h at room temperature. The band were detected using a Western Lighting Plus-ECL (PerkinElmer, Waltham, Massachusetts, USA) and visualized on X-ray film.
5
Pentamidine Effects on P21, P53 Expression
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Cells were cultured with vehicle or 2.5 μmol/L pentamidine in 6‐well plates (106 cells/well) at 37°C for 24 hours and then lysed using RIPA Lysis and Extraction Buffer (Thermo Scientific, 89901) containing the protease inhibitor cocktail (Thermo Scientific, 87786). Proteins were then separated by SDS‐polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. After blocked with 5% fat‐free milk for 1 hour at room temperature, the membranes were incubated with primary antibodies at 4°C overnight: P21 antibody (Proteintech, 10355‐1‐AP), P53 antibody (Proteintech, 10442‐1‐AP) and beta‐actin antibody (Proteintech, 7D2C10). The membranes were then incubated with horseradish peroxidase‐conjugated secondary antibodies for 1 hour at room temperature: Goat anti‐rabbit IgG (Proteintech, SA00001‐2) and goat anti‐mouse IgG (Proteintech, SA00001‐1).
6
Antibody Characterization Protocol
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DHA was obtained from Sigma-Aldrich (St. Louis, MO, USA). Cav1 (polyclonal, rabbit anti-human; cat. no. 16447-1-AP; dilution 1:1,000), MTCH2 (polyclonal, rabbit anti-human; cat. no. 16888-1-AP; dilution 1:1,000), β-tubulin (monoclonal, mouse anti-human; cat. no. 66240-1, dilution 1:2,000), β-actin (polyclonal, rabbit anti-human; cat. no. 23660-1-AP; dilution 1:1,000), GAPDH (polyclonal, rabbit anti-human; cat. no. 10494-1-AP; dilution 1:1,000) and Myc (monoclonal, mouse anti-human; cat. no. 60003-2-Ig; dilution 1:1,000) antibodies were obtained from ProteinTech Group, Inc. (Chicago, IL, USA); p53 antibody (monoclonal, mouse anti-human; cat. no. P8999; dilution 1:1,000) was obtained from Sigma-Aldrich; NAD(P)H:quinone oxidoreductase 1 (NQO1) antibody (monoclonal, mouse anti-human; cat. no. #3187, dilution 1:1,000) was obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA) and p84 antibody (monoclonal, mouse anti-human; cat. no. MA1-23261, dilution 1:1,000) was obtained from Invitrogen, Waltham, CA, USA. The secondary antibody (polyclonal, goat anti-rabbit/goat anti-mouse; cat. no. 042-06-15-06/042-06-18-06; dilution 1:10,000) was obtained from KPL, Inc. (Gaithersburg, MD, USA).
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