HL‐1 CM cells were seeded at a density of 1×105 cells/mL in 35‐mm dishes, and for a knockdown experiment, Snrk siRNA or control siRNA (Dharmacon) were transfected using Lipofectamine2000 reagent. Transfected cells were stimulated with and without Ang II (1 μmol/L) condition for 24 hours and were harvested using absolute methanol for 30 minutes, in 4°C to fix and permeabilize the cells. Cells were washed 3 times with fluorescence‐activated cell‐sorting buffer (1× PBS with 5% FBS and 0.1% NaN3) at 300g for 10 minutes and were subsequently incubated with anti–NF‐κB primary antibody (Cell Signaling Technologies, Danvers, MA; catalog No. 3033) per the manufacturer's protocol. After staining with primary antibody, the cells were washed 3 times and incubated with PE‐conjugated anti‐rabbit secondary antibody (1:500 dilution, Biolegend, San Diego, CA; catalog No. 406421) for 30 minutes at 4°C. Stained cells were washed 3 times and ran on a flow cytometer (BD LSR Fortessa, San Jose, CA). Sample acquisition was done using fluorescence‐activated cell‐sorting DIVA software (BD) and subsequently analyzed on FlowJo software (FlowJo, Ashland, OR). Secondary antibody controls were included for each corresponding experimental sample.
Anti nf κb primary antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti–NF‐κB primary antibody is a laboratory reagent used to detect and study the NF-κB protein in biological samples. It is a highly specific and sensitive tool for researchers investigating cellular signaling pathways and gene regulation processes.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Control sirna, by Horizon Discovery (1 mentions)
Diva software, by BD (1 mentions)
Chemiluminescent immunoblotting reagent, by Thermo Fisher Scientific (1 mentions)
Coomassie plus protein assay reagent, by Thermo Fisher Scientific (1 mentions)
Modified radioimmunoprecipitation assay ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)
Sensifast sybr lo rox kit, by Meridian Bioscience (1 mentions)
Anti asc primary antibody, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit or anti mouse igg, by Cell Signaling Technology (1 mentions)
Anti nlrp3 primary antibody, by Cell Signaling Technology (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Revertra ace qpcr master mix, by Toyobo (1 mentions)
Qiazol lysis reagent, by Qiagen (1 mentions)
2 protocols using anti nf κb primary antibody
1
Quantifying NF-κB Activation in Ang II-Stimulated Cells
2
Molecular Mechanisms of NLRP3 Inflammasome Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Dulbecco’s Modified Eagle Medium (DMEM) was purchased from Gibco (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from Thermo Scientific (Waltham, MA, USA). Modified radioimmunoprecipitation assay (RIPA) lysis buffer, protease inhibitor cocktail, Coomassie Plus™ Protein Assay Reagent, and chemiluminescent immunoblotting reagent were obtained from Thermo Fisher Scientific (Rockford, IL, USA). QIAzol lysis reagent was purchased from Qiagen (Valencia, CA, USA). ReverTra Ace® qPCR Master Mix was purchased from Toyobo Co., Ltd. (Osaka, Japan). SensiFASTTM SYBR® Lo-ROX Kit was purchased from Meridian Bioscience® (Cincinnati, OH, USA). The MTT or 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide dye and mouse anti-β-actin primary antibody were purchased from Sigma-Aldrich Company (St. Louis, MO, USA). Anti-NLRP3 primary antibody, anti-ASC primary antibody, anti-caspase-1 primary antibody, anti-NF-κB primary antibody, anti-PARP primary antibody, and goat horseradish peroxidase-conjugated anti-mouse- or anti-rabbit-IgG were obtained from Cell Signaling Technology company (Danvers, MA, USA).
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