The following antibodies and reagents were used: Sirtuin Antibody Sampler Kit #9787T, NF‐κB p65 Antibody Sampler Kit #4767, Death Receptor Antibody Sampler Kit #8356, antibodies against cleaved PARP #5626, cleaved caspase‐3 #9664, cleaved caspase‐8 #8592, cleaved caspase‐9 #9509, Bak #6947, Bax #5023, Bcl‐2 #15071, and Bcl‐X #2764 (Cell Signaling Technology, Danvers, MA, USA); antibodies against HDAC3 #sc‐130319 and β‐actin #sc‐58673 (Santa Cruz Biotechnology, Dallas, TX, USA); nicotinamide (NAM); and ammonium pyrrolidine dithiocarbamate (PDTC) (Sigma‐Aldrich, St Louis, MO, USA).
β actin sc 58673
Manufactured by Santa Cruz Biotechnology
Sourced in United States
β-actin (sc-58673) is a monoclonal antibody that specifically detects β-actin, a cytoskeletal protein found in eukaryotic cells. It is commonly used as a loading control or reference protein in Western blotting applications to normalize target protein expression levels.
Automatically generated - may contain errors
Lab products found in correlation
Cell culture supplements, by Thermo Fisher Scientific (2 mentions)
Bcl 2 15071, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3 9664, by Cell Signaling Technology (1 mentions)
Ammonium pyrrolidinedithiocarbamate pdtc, by Merck Group (1 mentions)
Cleaved caspase 9 9509, by Cell Signaling Technology (1 mentions)
Sirtuin antibody sampler kit, by Cell Signaling Technology (1 mentions)
Nf κb p65 antibody sampler kit, by Cell Signaling Technology (1 mentions)
Death receptor antibody sampler kit, by Cell Signaling Technology (1 mentions)
Bax 5023, by Cell Signaling Technology (1 mentions)
Nicotinamide, by Merck Group (1 mentions)
E cadherin 14472, by Cell Signaling Technology (1 mentions)
Hif 1α sc 13515, by Santa Cruz Biotechnology (1 mentions)
Bay 87 2243, by Selleck Chemicals (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Ab21624, by Abcam (1 mentions)
Ab97959, by Abcam (1 mentions)
P akt 4060 t, by Cell Signaling Technology (1 mentions)
Ecl reagent, by GE Healthcare (1 mentions)
Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions)
8 protocols using β actin sc 58673
1
Apoptosis Signaling Pathway Analysis
2
Investigating Cellular Signaling Pathways
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The MMP-2 (cat. no. 40994), MMP-9 (cat. no. 13667), t-JNK (cat. no. 9252), p-JNK (cat. no. 9255), anti-IgG antibodies (cat. no. 14708) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). The β-actin (sc-58673) and B cell lymphoma-2 (Bcl-2; sc-509) antibodies were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and the Sp1 antibody was purchased from Merck KGaA (Darmstadt, Germany). Secondary rat antibody conjugated with horseradish peroxidase (cat. no. A0208) was purchased from Biyuntian Biotechnology (Biyuntian, Beijing, China). Celecoxib (cat. no. S1261) and anisomycin (cat. no. S7409) were purchased from Selleck Chemicals (Houston, TX, USA). The small interfering (si)RNA for Sp1, 5′-UGUAGAGUCUGCCAACUGACCUGUC-3′, was purchased from Santa Cruz Biotechnology, Inc. The scramble siRNA, 5′-UAGUGCUUACGCAGUUGCUAGACCC-3′, was synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).
3
Melanoma and HEK293 Cell Culture
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WM35 and A375 melanoma cells and HEK293 cells were purchased from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) and authenticated and tested for contamination. They were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (Sijiqing, Hangzhou, China) at 37°C in a humidified atmosphere containing 5% CO2.
BAY 87-2243 (S7309) were purchased from Selleckchem. HIF1α (sc-13515, 1:1000), β actin (sc-58673, 1:1000) and GAPDH (sc-32233, 1:1000) were obtained from Santa Cruz Biotechnology. The antibodies recognizing E-Cadherin (#14472, 1:1000) and Vimentin (#3390, 1:1000) were purchased from Cell Signaling Technology.
BAY 87-2243 (S7309) were purchased from Selleckchem. HIF1α (sc-13515, 1:1000), β actin (sc-58673, 1:1000) and GAPDH (sc-32233, 1:1000) were obtained from Santa Cruz Biotechnology. The antibodies recognizing E-Cadherin (#14472, 1:1000) and Vimentin (#3390, 1:1000) were purchased from Cell Signaling Technology.
4
Western Blotting Technique for Protein Analysis
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Western blotting was carried out as described previously [24 (link)]. In brief, for Western blotting, proteins were extracted using RIPA buffer mixed with protease inhibitors (1:100), phenylmethylsulfonylfluoride (PMSF, 1:100) and dithiothreitol (DTT, 1:100) at 4 °C for 30 min. The lysate was centrifuged at 13000 rpm at 4 °C for 15 min. The supernatants were collected and protein concentrations were measured using the BCA method. For Western blot analysis, protein samples were separated by 8% SDS-PAGE and transferred onto polyvinylidene difluoride membranes (Bio-Rad). The membranes were then blocked with 5% nonfat milk and incubated with primary antibodies overnight followed by washing and reaction with horseradish peroxidase–conjugated secondary antibody for 1 h at 37 °C. The reaction was detected by using ECL reagents (GE Healthcare) and the signals were captured by X-ray films. Antibodies against RIF1 (sc-515,573) and β-actin (sc-58,673) were obtained from Santa Cruz Biotechnology. Antibodies against NANOG (ab21624) and SOX2 (ab97959) were purchased from Abcam. Antibodies against PI3K (4257 T), p-PI3K (4228 T), AKT (4691 T) and p-AKT (4060 T) were purchased from Cell Signaling Technology. Antibody to hTERT (AP1410d) was purchased from Abgent.
5
Investigating miR-650 Regulatory Mechanisms
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MTT was purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Four synthetic, chemically modified short double-stranded RNA oligonucleotides (miR-650 mimics, mimics negative control, miR-650 inhibitor and inhibitor negative control) were obtained from Guangzhou RiboBio Co., Ltd. (Guangzhou, China). Primary antibodies against p38 (#9212), phosphorylated (p)-p38 (#4511), ERK1/2 (#4695), p-ERK1/2 (#4370) and RhoA (#2117) were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA); active Rac1 (#26903) and active RhoA (#26904) from NewEast Biosciences (King of Prussia, PA, USA); E-cadherin (ab1416), β-catenin (ab32572), Rac1 (ab33186) and FITC-conjugated anti-rat (ab6717)/anti-mouse IgG (ab6785) secondary antibodies from Abcam (Cambridge, UK); α-smooth muscle actin (α-SMA, 55135–1-AP) from ProteinTech Group, Inc. (Chicago, IL, USA); vimentin (BM0135) from Wuhan Boster Biological Technology, Ltd. (Wuhan, China); and β-actin (sc-58673) and HRP conjugated goat anti-rabbit/goat anti-mouse secondary antibodies (sc-2004/sc-2005) from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The primary antibody ING4 (#40-7700) and the fluorescent dye Alexa Fluor 546 conjugated phalloidin (A22283, 1:5,000) were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA).
6
Investigating Inflammatory Signaling Pathways
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Cell culture supplements were purchased from Invitrogen (Carlsbad, CA, USA). Antibodies against IL-6 (SC-28343), p-p65 (Ser536; SC-101752), p-65 (SC-8008), p-85 (SC-1637), p-AKT (Thr308; SC-16646-R), AKT (SC-5298), p-IKKα/β (Ser180/Ser181; SC-23470-R), IKKα/β(SC-7607), p-IκBα (Ser32; SC-8404), IκBα (SC-203), PCNA(SC-36), and β-actin (SC-58673), and C6 ceramide (an ERK activator), were all purchased from Santa Cruz (Santa Cruz, CA, USA). Anti-TNF-α (a11534) was obtained from Abclonal (Woburn, MA, USA). Antibody against p-p85 (Tyr458/Tyr199; 4228S) was purchased from Cell Signaling (Danvers, MA, USA). AKT inhibitor were supplied by Calbiochem (San Diego, CA, USA). Ly294002, TPCK and PDTC were bought from Enzo Life Sciences International (Plymouth Meeting, PA, USA) and Cell culture supplements were purchased from Invitrogen (Carlsbad, CA, USA). Dual-Luciferase® Reporter Assay System was bought from Promega (Madison, WI, USA). All other chemicals not described above were supplied by Sigma-Aldrich (St Louis, MO, USA).
7
Investigating Inflammatory Signaling Pathways
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Antibodies against TNF-α (A11534), IL-1β (SC-7884), IL-8 (ab18672), p-FAK (Thr202/Tyr204; SC-7383), FAK (SC-5298), p-85 (SC-1637), p-AKT (Thr308; SC-16646-R), AKT (SC-5298), p-p65 (Ser536; SC-101752), p-65 (SC-8008), and β-actin (SC-58673) were purchased from Santa Cruz (Santa Cruz, CA, USA). Activators Angiotensin II (a FAK activator), 740-YP (a PI3K activator), SC-79 (an AKT activator) and prostratin (an NF-κB activator) were all purchased from Santa Cruz (Santa Cruz, CA, USA). Cell culture supplements were purchased from Invitrogen (Carlsbad, CA, USA). A Human Inflammation Antibody Array kit was supplied by RayBiotech, Inc. Human TNF-α, IL-1β and IL-8 ELISA kits were purchased from R&D Systems (MN, USA).
8
Western Blot Analysis of VEGF-C
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Clinical tissues or cultured cells were lysed with RIPA buffer (Beyotime Biotechnology, China). Equal amounts of proteins were separated by 10% polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto Immobilon-P Transfer membrane (Merck Millipore, China). Immunoblots were prepared using standard protocols including BSA blocking, primary antibody incubation (VEGF-C, sc-374628; β-actin, sc-58673; both from Santa Cruz Biotechnology, USA), secondary horseradish peroxidase-conjugated antibody incubation (anti-goat IgG for VEGF-C, anti-mouse IgG for TBL1XR1 and β-actin) and eventually developed with chemiluminescence reagents (Santa Cruz Biotechnology, USA).
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