Vitamin C (Tianjin Kingyork, H12020392), Vitamin D3 (Beijing Solarbio Science & Technology, V8070), Mouse TNF-alpha ELISA Kit (RayBiotech, P06804), Mouse IL-6 ELISA Kit (RayBiotech, P08505), Mouse IL-1β ELISA Kit (Shanghai Tongwei Biotechnology, TW56961), Mouse insulin ELISA Kit (Shanghai Tongwei Biotechnology, TW085566), Mouse endotoxin (ET) ELISA Kit (Jiangsu Meimian, MM-0369M1), Total Cholestenone Content Assay Kit (Beijing Solarbio Science & Technology, BC 1985), Triglyceride Content Assay Kit (Beijing Solarbio Science & Technology, BC0625), Total SOD activity detection kit (Shanghai Beyotime Biotechnology, S0101S), Lipid oxidation detection kit (Shanghai Beyotime Biotechnology, S0131S), BCA protein concentration assay kit (Shanghai Beyotime Biotechnology, P0010), Oil red O (Sigma, O0625), Anti-FXR1 antibody (Abcam, ab155124), BSEP Monoclonal antibody (Proteintech Group, 67512-1-Ig), Anti-Fas antibody (Abcam, ab271016), Anti-ZO1 tight junction protein antibody (Abcam, ab221547), Anti-Occludin antibody (Abcam, ab222691), Anti-SLC10A2/ASBT antibody (Abcam, ab203205), TRIzol Reagent (Thermo Fisher, 15596026), cDNA synthesis kit (Beijing Tiangen Biotech, KR118), and SuperReal PreMix Plus (SYBR Green) (Beijing Tiangen Biotech, FP205) were used.
Anti fas antibody
Manufactured by Abcam
Sourced in United States, Denmark
Anti-Fas antibody is a laboratory reagent used for the detection and quantification of the Fas protein, also known as CD95 or Apo-1. Fas is a cell surface receptor that plays a role in programmed cell death (apoptosis). The antibody can be used in various immunological techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and localization of Fas in biological samples.
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Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Vitamin d3, by Solarbio (1 mentions)
Mouse tnf alpha elisa kit, by RayBiotech (1 mentions)
Mouse il 6 elisa kit, by RayBiotech (1 mentions)
Total cholestenone content assay kit, by Solarbio (1 mentions)
Total sod activity detection kit, by Beyotime (1 mentions)
Anti zo1 tight junction protein antibody, by Abcam (1 mentions)
Anti occludin antibody, by Abcam (1 mentions)
Triglyceride content assay kit, by Solarbio (1 mentions)
Oil red o, by Merck Group (1 mentions)
Superreal premix plus sybr green, by Tiangen Biotech (1 mentions)
Bca protein concentration assay kit, by Beyotime (1 mentions)
Cdna synthesis kit, by Tiangen Biotech (1 mentions)
Lysis buffer, by Merck Group (1 mentions)
Nitrocellulose membrane, by Cytiva (1 mentions)
Anti t bet tbx21antibody, by Abcam (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Echomri 3 in 1 instrument, by Echo Medical Systems (1 mentions)
Anti actin antibody, by Merck Group (1 mentions)
Quantikine elisa, by R&D Systems (1 mentions)
6 protocols using anti fas antibody
1
Investigating Liver Inflammation and Metabolism
2
Immunoblot Analysis of Immune Signaling
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BMCs (1 × 107) from each mouse were lysed in 0.5 mL of lysis buffer (Sigma, St. Louis, MO, USA). The extracts were cleared by centrifuging at 10,000g and 4°C for 15 min and diluted with the lysis buffer to achieve about 2 mg/mL protein concentration. Protein samples were separated on 10% SDS-PAGE and transferred onto nitrocellulose membranes (Amersham Pharmacia Biotech, Uppsala, Sweden). The membranes were incubated with primary antibodies, including anti-T-bet/Tbx21antibody (1 : 1000), anti-Fas antibody (1 : 1000) (Abcam, Cambridge, MA, USA), and anti-mouse-caspase-3, anti-mouse-cleaved caspase-3 antibody, anti-mouse-eukaryotic initiation factor 2 (eIF2) α (D7D3), anti-mouse-phospho-eIF2α (Ser51) (D9G8), anti-mouse-Fyn, anti-mouse interferon regulatory factor-1 (IRF-1), anti-mouse-signal transducer and activator of transcription (Stat) 1, and anti-mouse-Stat3 rabbit monoclonal antibodies (1 : 1000, CST, Boston, MA, USA) and incubated with horseradish peroxidase-conjugated secondary antibody (CST). All immunoreactive proteins were visualized with SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific, Rockford, IL, USA). Densitometry plots showing protein expression were normalized to GAPDH.
3
Metabolic Profiling in Fasted Mice
4
Immunohistochemical Analysis of Fas Protein
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Formalin-fixed paraffin-embedded cells were deparaffinized in xylene and rehydrated in graded concentrations of ethanol. For antigen retrieval, the sections were treated in a micro-oven for 10 min in citrate buffer (pH 9). After cooling, slides were washed in wash buffer (Tris-buffered saline [TBS] + 0.1% Triton X) and covered with cover plates following incubation with primary antibody (anti-Fas antibody 1:1,500, Abcam, Denmark) overnight at 4°C. On the next day, slides were washed with wash buffer, incubated with the HiDef detection HRP Polymer System (AH Diagnostics, Denmark) for 20 min at 4°C, and developed using Dab+ (Dako, Denmark). Slides were dehydrated, covered with coverslips, and viewed under a microscope for quantification.
5
Apoptosis Signaling Pathway Analysis
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PPTT-treated cells were lysed in 0.3 mL of lysis buffer (50 mM Tris–HCl (pH 8.0), 150 mM NaCl, 1% TritionX-100, 100 μg mL–1 PMSF). The cell lysates were centrifuged at 12 000 rpm for 20 min and then collected as the supernatant. Equal amounts of proteins were separated using 10–15% SDS-PAGE gel and blotted onto a polyvinylidene difluoride membrane. The membrane was first incubated with primary antibodies (1 : 1000 v/v) overnight at 4 °C. The primary antibodies used were anti-Fas antibody (Abcam) and rabbit polyclonal BID (Cell Signaling Technology). The membrane was rinsed and then incubated for 1 h with peroxidase conjugated secondary antibodies (1 : 10 000, Abcam). Chemiluminescence detection was performed with an ECL kit (KGP1201) from KeyGen Biotech (China).
6
Immunohistochemical Detection of Fas Receptor
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Tissues were fixed in 4% neutral buffered formalin, processed, then embedded in paraffin and cut into 5-mm sections. Tissue sections were deparaffinized and rehydrated. Endogenous peroxidase was blocked in 3% H 2 O 2 in phosphate-buffered saline for 10 minutes. After blocking nonspecific sites with 1.5% blocking serum in phosphatebuffered saline for 1 hour at room temperature, tissue sections were incubated for 1 hour at room temperature with the anti-Fas antibody (1:50 dilution; Abcam). After a 30-minute reaction with a biotinylated secondary antibody, slides were washed with phosphate-buffered saline and incubated with streptavidin conjugated with horseradish peroxidase for 10 minutes. The reaction was then revealed with diaminobenzidine. The slides were mounted with Eukitt and observed with an Olympus BX60 microscope (Olympus, Center Valley, PA). Images were captured with IPE software version 6.0 (Aspen Tech, Bedford, MA).
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