The cells were lysed using sodium dodecyl sulfate buffer containing proteinase inhibitors (Roche). Equal amounts of protein (50 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride membranes (Bio-Rad Laboratories, Shanghai, China). The membranes were blocked and incubated with Antibodies against GAPDH (dilution 1:200, Cat. no. sc-47724), IL-6 (1:200, Cat. no. sc-65327), p53 (1:200, Cat. no. sc-47698) and IL-17R (1:100, Cat. no. sc-376374) overnight at 4°C. Antibodies against GAPDH, IL-6, p53 and IL-17R were purchased from Santa Cruz Biotechnology. The membranes were then incubated with horseradish peroxidase-labeled secondary antibody (1:500, Cat. no. sc-2031) (Santa Cruz Biotechnology). The protein bands were visualized using an enhanced chemiluminescence reagent.
Antibodies against
Manufactured by Santa Cruz Biotechnology
Sourced in United States, Canada
Antibodies against are a type of laboratory reagent used to identify and detect specific proteins or molecules in biological samples. These antibodies are produced by the immune system and can be isolated and used for various research and diagnostic applications. The core function of antibodies against is to bind to and recognize their target analyte, enabling researchers to study its presence, distribution, and interactions within a sample.
Automatically generated - may contain errors
Lab products found in correlation
Antibodies against, by Cell Signaling Technology (6 mentions)
Bca kit, by Thermo Fisher Scientific (6 mentions)
Gapdh, by Santa Cruz Biotechnology (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Caspase 3, by Cell Signaling Technology (3 mentions)
β actin, by Santa Cruz Biotechnology (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Smartpool sirna, by Horizon Discovery (2 mentions)
Antibodies specific, by Abcam (2 mentions)
Signalsilence sirna, by Cell Signaling Technology (2 mentions)
Antibody against phospho ser, by Meridian Bioscience (2 mentions)
Fabppm got2, by GeneTex (2 mentions)
3 utr specific duplex sirnas, by OriGene (2 mentions)
Slc27a1, by Horizon Discovery (2 mentions)
3 utr specific trilencer sirna targeting, by OriGene (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Cell culture medium, by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Rankl, by R&D Systems (2 mentions)
63 protocols using antibodies against
1
Western Blot Analysis of Cellular Proteins
2
Arctigenin Mechanism of Action
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Arctigenin was purchased from Enzo Life Sciences (Farmingdale, NY, USA). The chemical structure of arctigenin is as shown in Fig. 1A . All reagents used for cell culture were purchased from Thermo Scientific (Fremont, CA, USA). Antibodies against HO-1, Nrf-2, c-Jun, and lamin A were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Antibodies against phospho- or total forms of ERK, JNK, p38, and AKT were obtained from Cell Signaling Technology (Danvers, MA, USA). All reagent used for RT-PCR were purchased from Promega (Madison, WI, USA).
3
Sirtinol-mediated Regulation of FoxO3a
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Sirtinol was purchased from Calbiochem (Darmstadt, Germany). Sirtinol was dissolved in 100% DMSO at concentration of 10 mM and stored at −20°C until use. The following compounds were obtained from Gibco BRL (Maryland, USA): Dulbecco's modified eagle medium (DMEM), Ham's F-12 Nutrient Mixture (F-12) fetal bovine serum (FBS), trypan blue, penicillin G, and streptomycin. Dimethyl sulfoxide (DMSO), ribonuclease A (RNase A), and propidium iodide (PI) were purchased from Sigma-Aldrich (Missouri, USA). Antibodies against FoxO3a were obtained from Epitomics (California, USA). Annexin V-FITC staining kit was purchased from Strong Biotech (Taipei, Taiwan). Antibodies against phospho-Akt, β-catenin, Sirt1, and β-actin were purchased from Santa Cruz Biotechnology (California, USA). Anti-mouse and anti-rabbit IgG peroxidase-conjugated secondary antibodies were purchased from Pierce (Illinois, USA). The anti-rabbit Rhodamine-conjugated antibody was purchased from Abcam (Cambridge, UK).
4
Comprehensive Biochemical Assay Protocol
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Folin–Ciocalteu reagent, silver nitrate and gold chloride solution, as well as Bradford reagent and 2-thiobarbituric acid were purchased from Sigma-Aldrich Chemicals GmbH (Darmstadt, Germany). Kits for capase-3 and capase-8 determinations were purchased from Elabscience (Houston, TX, USA). Antibodies against p53, BCL2, BAX, NFkB, pNFkB proteins and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against Ki-67 were purchased from Invitrogen (Rockford, IL, USA) and those against PCNA and MAPK were purchased from Cell Signaling Technology (Danvers, MA, USA). Propidium iodide and Annexin V-fluorescein isothiocyanate were purchased from BD Pharmingen Biosciences (San Jose, CA, USA). The CellTiter 96® cell proliferation assay was purchased from Promega Corporation (Madison, WI, USA). All reagents used were of high purity.
5
Antibody generation and siRNA targeting
6
Western Blot Analysis of Iron Regulation
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Western blot was performed as described (Zhang et al. 2014 (link)). Antibodies against p53, p21, IRP2, and TfR1 were purchased from Santa Cruz Biotechnology. Anti-mouse p53 (1C12) was purchased from Cell Signaling Technology. Antibodies against FDXR, FDX1, FDX2, IRP1, and FTH1 were purchased from Abcam. Anti-HA was purchased from Covance. Anti-actin and HRP-conjugated secondary Antibodies against rabbit or mouse IgG were purchased from Bio-Rad. The immunoreactive bands were visualized by enhanced chemoluminescence (Thermo Fisher Scientific, Inc.) and quantified by densitometry with the BioSpectrum 810 imaging system (UVP LLC).
7
Antibody Characterization of ATPase Subunits
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Chemicals of the highest purity and culture media were purchased from Sigma and Invitrogen. Akt and phosphorylated (Ser473) Akt anitbodies were purchased from Cell Signaling Technology (Danvers, MA). Antibodies against ERK 1/2, phosphorylated ERK 1/2, α2 subunit of Na+/K+-ATPase, PI3K p110α, PI3K p110γ, PI3K p85α, Src, goat anti-mouse IgG-horseradish peroxidase, and goat anti-rabbit IgG-horseradish peroxidase were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibody against Na+/K+-ATPase α1 subunit (α6F) was purchased from Developmental Studies Hybridoma Bank (Iowa City, IA). Na+/K+-ATPase α3 subunit antibodies (MA3-915) were purchased from Pierce Biotechnology (Rockford, IL).
8
Protein and Gene Expression Analysis
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Western blot and real-time PCR analysis were performed as described previously [17 (link)]. Antibodies against NFATc1 and actin were purchased from Santa Cruz Biotechnology (CA, USA). Primers (Table 1 ) were chosen with the online Primer3 design program [32 (link)].
9
Osteoclastogenesis Assay with M-CSF and RANKL
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Penicillin, streptomycin, cell culture medium, and fetal bovine serum (FBS) were purchased from Invitrogen Life Technologies. Mouse soluble macrophage-colony stimulating factor (M-CSF) and RANKL were purchased from R&D Systems. The CCK-8 assay kit was purchased from Dojindo Molecular Technologies. Antibodies against nuclear factor of activated T cells (NFAT)c1, c-Fos, and actin were purchased from Santa Cruz Biotechnology and Antibodies against MAP kinases from Cell Signaling Technology. Matairesinol was purchased from Sigma-Aldrich and dissolved in DMSO (dimethylsulfoxide; Sigma-Aldrich).
10
Osteoclastogenesis Signaling Pathway Profiling
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Antibodies against phospho-Jun N-terminal kinase (p-JNK), JNK, phospho-p38, p38, phosphor-extracellular signal-regulated kinase (p-ERK), ERK, IκBα, and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against c-Fos and NFATc1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Recombinant macrophage colony-stimulating factor (M-CSF) and RANKL were obtained as previously described [11 (link)].
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