ASP, Hoechst 33258 and sodium hypochlorite (HClO) solution were purchased from Sigma‐Aldrich Co. (St. Louis, MO, USA). Commercial enzyme‐linked immunosorbent assay (ELISA) kits were supplied by Boster BioTech Co. (Wuhan, China) for measuring MPO, interleukin (IL)‐6 and tumour necrosis factor (TNF)‐α. Primary antibody against COX‐2 or CSE was purchased from Bioworld Technology Inc. (Louis Park, MN, USA). β‐Actin was used as a loading control, whose primary antibody was purchased from KangChen Bio‐tech Inc. (Shanghai, China). Cell counting kit (CCK)‐8 and rhodamine (Rh)123 were purchased from Dojindo Laboratory (Kyushu, Japan). DMEM and foetal bovine serum (FBS) were supplied by Gibico BRL (Ground Island, NY, USA). Other compounds were provided by Thermo Fisher Scientific Inc. (Shanghai, China).
β actin
Manufactured by Kangchen Biotech
Sourced in China
β-actin is a highly conserved cytoskeletal protein that is essential for maintaining the structural integrity of eukaryotic cells. It is commonly used as a housekeeping gene for normalization in gene expression analysis.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33258, by Merck Group (1 mentions)
Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
P mtor ser2448, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Mdivi 1, by Merck Group (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Cleaved caspase 9, by Cell Signaling Technology (1 mentions)
Luteolin, by Santa Cruz Biotechnology (1 mentions)
Bnip3, by Cell Signaling Technology (1 mentions)
Lamp1, by Abcam (1 mentions)
P drp1 ser616, by Cell Signaling Technology (1 mentions)
Pink1, by Abcam (1 mentions)
Vinculin, by Abcam (1 mentions)
Doxorubicin, by Beyotime (1 mentions)
Parkin, by Cell Signaling Technology (1 mentions)
Rabbit anti tbx5 polyclonal antibody, by Abcam (1 mentions)
P egfr, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
6 protocols using β actin
1
Assessing Anti-Inflammatory Potential
2
Protein Extraction and Western Blot
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Cells were collected and washed twice with PBS, and protein was extracted by whole-cell lysis with a kit purchased from Beyotime (Haimen, Jiangsu, China) that contained protease and phosphatase inhibitors. Centrifugation at 4 °C removed cellular debris, and the protein concentration was determined by a Pierce BCA assay. The protein content was electrophoresed on a 10% SDS-PAGE gel and then immunoblotted on a polyvinylidene fluoride membrane (American Biosciences). Antibodies against PARP (9532S, Cell Signaling Technology), PCNA (2586, Cell Signaling Technology), Caspase-9 (9508S, Cell Signaling Technology), and β-actin (KC-5A08, Kangchen Biotech) were used.
3
Hippocampal Protein Quantification
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Protein in the bilateral hippocampus was extracted. Bicinchoninic acid was used for quantitative analysis. Electrophoresis was done with 10% polyacrylamide gel. Transmembrane was used with polyvinylidene fluoride (PVDF). The membrane was blocked with TTBS (NaCl 150 mM, Tris 25 mM, pH 7.6), and then probed with β-actin (1:500 in blocking solution for 14–16 hours at 4°C, Kangchen Bio-tech, Shanghai, China). The secondary antibody IgG (Cell Signaling, USA) was applied for two hours with slow shaking at room temperature. After each incubation, the membrane was thoroughly washed with TTBS, and then the membrane was treated with an ECL kit (Pierce, USA) and by Western blot.
4
Mitochondrial Dynamics in Cancer Cell Apoptosis
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Doxorubicin (Beyotime Institute of Biotechnology, Shanghai, China), luteolin (≥98%, Santa Cruz Biotechnology, sc-203119), mdivi-1 (≥98%, Sigma Aldrich, M0199). Bax (1:1,000, Cell Signaling Technology, #5023S), Bcl-2 (1:1,000, Cell Signaling Technology, #15071S), Bnip3 (1:1,000, Cell Signaling Technology, #44060), cleaved caspase-9 (1:1,000, Cell Signaling Technology, #9509S), Drp1 (1:1,000, Cell Signaling Technology, #8570), p-Drp1 (Ser616) (1:1,000, Cell Signaling Technology, #3455S), LAMP1 (1:500, Abcam, ab208943), LC3B (1:1,000, Abcam, ab48394), mTOR (1:1,000, Cell Signaling Technology, #2983S), p-mTOR (Ser2448) (1:1,000, Cell Signaling Technology, #5536S), P62 (1:1,000, Cell Signaling Technology, #5114S), parkin (1:1,000, Cell Signaling Technology, #4211), Pink1 (1:1,000, Abcam, ab216144), TFEB (1:500, Cell Signaling Technology, #32361S), vinculin (1:1,000, Abcam, ab129002);anti-mouse Alexa Fluor (1:1000, Cell Signaling Technology, #4408), anti-rabbit Alexa Fluor (1:1,000, Cell Signaling Technology, #8890);β-actin (1:5,000, KangChen Bio-tech, Shanghai, China).
5
TBX5 Protein and mRNA Expression Regulation
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HEK 293T cells were transfected with 2 μg of miRNA expression vector or 100 nm of miRNA inhibitors in 6-well culture plates. Cells were harvested and lysed 48 h after transfection in RIPA buffer and assayed for the relative protein concentrations of intracellular TBX5 (rabbit anti-TBX5 polyclonal antibody; Abcam) and β-actin (Kangchen Bio-tech) using western blot analysis. RNA was also purified from these cells for quantitative RT-PCR analysis of TBX5 mRNA levels.
6
RCC Cell Lines and Reagents
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Human RCC cell lines 769-P, 786-O, ACHN were obtained from the American Type Culture Collection (ATCC). Human RCC cell lines Caki-1, OS-RC-2 were purchased from National Platform of Experiment Cell Resources for SciTech (Beijing, China), and Human RCC cell lines RCC42 was kindly presented by Dr. JerTsong Hsieh (University of Texas Southwestern Medical Center, Dallas, TX, USA). All cell lines were maintained in RPMI1640 medium with 10% fetal bovine serum (FBS) at 37 ˚C and 5% CO2. Cells used in experiments were in good condition without mycoplasma contamination. LY294002 was obtained from Selleckchem (Houston, TX, USA). The antibodied were as follows: MUC15 (Rabbit, Sigma-Aldrich, St Louis, MO, USA), β-actin and GAPDH (mouse, Kangchen Bio-tech, Shanghai, China), AKT, p-AKT, p-EGFR, p-GSK-3β, β-Catenin and MMP2 (Rabbit, Cell Signaling Technology, Danvers, MA, USA), MMP9 (Rabbit, abcam, Inc., Cambridge, Britain).
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