In this study, sucrose (Amresco; 20140319) was employed in the sucrose consumption test, and fluoxetine (Eli Lilly and Company, Suzhou 215021, China; 4126A) was used as the standard control for acupuncture. ELISA Kit of BDNF (BlueGene Biotech CO., LTD., China; E02B0029) was used to detect the expression of BDNF in serum. The antibodies and some reagents used for WB analysis included rabbit polyclonal anti-BDNF (LifeSpan BioSciences; LS-C343943), rabbit polyclonal anti-acH3K9 (Cell Signaling Technology; 9649S), mouse anti-HDAC2 (Cell Signaling Technology; 5113S), β-actin (Zhongshanjinqiao Biotechnology Co., Ltd., Beijing, China; TA-09), goat anti-rabbit IgG (Jackson; 111-035-003), rabbit anti-H3 (Cell Signaling Technology, 4499S), ECL (Millipore; WBKLS0500), and bicinchoninic acid (BCA) protein assay kit (Cwbiotech; 02912E). Furthermore, the reagents used for RT-PCR and methylation-specific quantitative real-time PCR analysis included ultrapure RNA kit (Cwbiotech), HiFi-MMLV Reverse Transcriptase (Cwbiotech), SYBR Green PCR Mixture (Cwbiotech), and Dnase I (Cwbiotech), etc.
β actin
Manufactured by Zhongshan Jinqiao Biotechnology
Sourced in China
β-actin is a highly conserved cytoskeletal protein that plays a fundamental role in cell structure and function. It is commonly used as a reference gene or loading control in various molecular biology techniques, such as Western blotting, immunohistochemistry, and quantitative PCR, to normalize and compare gene expression levels across samples.
Automatically generated - may contain errors
Lab products found in correlation
Sucrose, by Avantor (1 mentions)
Rabbit anti h3, by Cell Signaling Technology (1 mentions)
Dnase 1, by CWBIO (1 mentions)
Anti ach3k9, by Cell Signaling Technology (1 mentions)
Mouse anti hdac2, by Cell Signaling Technology (1 mentions)
Hifi mmlv reverse transcriptase, by CWBIO (1 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
Fluoxetine, by Eli Lilly (1 mentions)
Ultrapure rna kit, by CWBIO (1 mentions)
Goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Bca protein assay kit, by CWBIO (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
T per tissue protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Zhongshan Jinqiao Biotechnology (1 mentions)
Phospho p65, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
3 3 diaminobenzidine (dab), by Zhongshan Jinqiao Biotechnology (1 mentions)
Nf κb p65, by Zhongshan Jinqiao Biotechnology (1 mentions)
Bcl 2, by Zhongshan Jinqiao Biotechnology (1 mentions)
6 protocols using β actin
1
Sucrose Consumption and Fluoxetine in BDNF
2
Spleen Protein Extraction and Western Blot Analysis
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Spleen tissues were homogenized in T-PER Tissue Protein Extraction Reagent (Thermo Fisher Scientific, Inc.) and the proteins were extracted. The protein concentrations in the samples were determined using a bicinchoninic acid protein assay kit. Equal amounts of proteins (30 µg) from homogenates were resolved using 10% SDS-PAGE prior to being transferred to nitrocellulose membranes. Membranes were blocked in TBS containing 0.1% Tween-20 (TBST) and 5% nonfat dry milk for 2 h at room temperature. Following washing with TBST, the membranes were incubated with primary antibodies for CD20 (dilution, 1:300; Santa Cruz Biotechnology, Inc., cat. no. SC-7735) or IL-10 (dilution, 1:1,000; Abcam; cat. no. Ab189392) at 4°C overnight. Then the membranes were washed again and incubated with the appropriate horseradish peroxidase-conjugated secondary antibodies (dilution, 1:3,000; Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., cat. nos. ZB-2306 and ZB-2305) for 1 h at room temperature. The membrane was further washed with TBST for 30 min and incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, Inc.). β-actin (dilution, 1:1,000; Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., cat. no. TA-09) was used as an internal control. Immunoreactive proteins were analyzed using ImageJ v1.48 (National Institutes of Health, Bethesda, MA, USA).
3
Western Blot Analysis of Cell Signaling
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Cells were lysed in RIPA buffer with protease inhibitors and phosphatase inhibitors [15 (link)–16 (link)]. The protein was resolved by SDS/PAGE and blotted on PVDF membranes (Millipore, Bedford, MA, USA)[15 (link)–16 (link)]. The PVDF membranes were incubated with specific primary antibodies at 4°C overnight [15 (link)–16 (link)]. After incubation with HRP-linked secondary antibodies, immunoreactive proteins were visualized using DAB (Beijing Zhongshan Jinqiao biotechnology Co., Ltd.).
Primary antibody against GRα was from Santa Cruz Biotechnology. IκBα, phospho-p65 and cleaved caspase 3 were from Cell Signaling Technology (Beverly, MA, USA). NF-κB p65, Bcl-2, β-actin and HRP-linked secondary antibodies were from Beijing Zhongshan Jinqiao biotechnology Co., Ltd.
Primary antibody against GRα was from Santa Cruz Biotechnology. IκBα, phospho-p65 and cleaved caspase 3 were from Cell Signaling Technology (Beverly, MA, USA). NF-κB p65, Bcl-2, β-actin and HRP-linked secondary antibodies were from Beijing Zhongshan Jinqiao biotechnology Co., Ltd.
4
Western Blot Analysis of CXCR1 and CXCR2 in Liver Cancer Cells
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In brief, liver cancer cells were lysed using radioimmunoprecipitation assay buffer (BIOSS) for 30 min on ice. The cell lysate was centrifuged at 12,000 × g for 20 min at 4°C and the supernatant was collected. Protein concentration was determined by BCA protein assay. A total of 30 µg protein/lane was separated by SDS-PAGE on a 10% gel and transferred onto nitrocellulose filter membranes. Following blocking with 5% skim milk for 1 h at room temperature, the membranes were incubated with primary antibodies overnight at 4°C, and then with secondary antibodies for 2 h at room temperature. The primary antibodies used were as follows: Rabbit polyclonal anti-CXCR1 (1:2,000; cat. no. ab137351; Abcam) and anti-CXCR2 (1:2,000; cat. no. ab14935; Abcam) antibodies; monoclonal mouse anti-GAPDH (1:2,000; cat. no. TA-08; Zhongshan Jinqiao Biotechnology) and β-actin (1:2,000, cat. no. TA-09; Zhongshan Jinqiao Biotechnology, Beijing, China). The goat anti-mouse (cat. no. ZB2305; 1:10,000) and anti-rabbit secondary antibodies (cat. no. ZB2301; 1:10,000) were purchased from Zhongshan Jinqiao Biotechnology. The protein bands were visualized by ECL reagents (cat. no. C05-07003; BIOSS). Images were captured and the intensity of the bands was quantitated with the Bio-Rad Versa Doc imaging system (Bio-Rad Laboratories, Inc.).
5
Crocetin and ATO Antioxidant Mechanism
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Crocetin with HPLC purity of ≥ 98% and arsenic trioxide (ATO, MW:197.84) were procured from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD) detection kits were procured from Jiancheng Bioengineering Institute of Nanjing (Nanjing, Jiangsu, China). The ROS detection kit was procured from Wuhan Servicebio Technology, Co., Ltd (Wuhan, China). Antibodies specific for Nrf2 and NQO1 were procured from Bioworld Technology, Co., Ltd. (Nanjing, Jiangsu, China). HO-1 antibodies were obtained from Wuhan Sanying Biotechnology Co., Ltd (Wuhan, China). β-actin was procured from Beijing Zhongshan Jinqiao Biotechnology Co., Ltd (Beijing, China). The IL-6, IL-1β, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Wuhan Servicebio Technology CO., Ltd (Wuhan, China).
6
Western Blot Analysis of Cell Signaling Proteins
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Western blot analysis was performed as described in the literature [27 (link)]. In brief, the total proteins in cells were extracted using cell lysate, and were quantified. The extracted proteins were separated by 10–15% SDS-PAGE and transferred to cellulose acetate membrane. The 5% skim milk powder solution was used to close the membrane for 2 h, then the primary antibody was used to incubate the membrane at 4 °C overnight. The goat IgG with HRP conjugate (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was incubated at room temperature for 1 h. The protein band was then colored and the grayscale value of the protein band was analyzed using ImageJ software. The β-actin (1, 5000, Beijing zhongshan jinqiao biotechnology co., LTD., Beijing, China) was used as an internal control. Among them, mTOR, Akt, PI3K I, Bcl-2, PI3K III, Beclin-1 and LC3 were obtained from Abcam (Cambridge, Massachusetts, USA) with a dilution ratio of 1: 1000.
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