Cell lysates were prepared using ice-cold RIPA lysis buffer (50 mM Tris-Cl, 150 mM NaCl, 1% NP-40, 1% Triton X-100) supplemented with 1% protease inhibitor cocktail (Sigma-Aldrich). After centrifugation at 14 000g at 4 °C for 20 min, the resulting supernatant was collected and subject to the total protein assay using Protein Assay Dye Reagent Concentrate (Bio-Rad). Proteins were resolved by 7.5% SDS-PAGE and transferred onto PVDF membranes (Bio-Rad) via semidry transfer. After blocking in 5% nonfat dry milk, membranes were incubated with primary antibodies (anti-P-gp (Abcam) or anti-β-actin (Enzo)) at 4 °C overnight. Membranes were washed and incubated with appropriate peroxidase-conjugated secondary antibodies for 1 h at room temperature. Proteins were visualized on Kodak BioMax XAR Films (Sigma-Aldrich) using ECL.
Anti β actin
Manufactured by Enzo Life Sciences
Sourced in United States
Anti-β-actin is a primary antibody that specifically recognizes and binds to the β-actin protein, a cytoskeletal protein found in eukaryotic cells. It can be used in various immunological techniques, such as Western blotting, immunocytochemistry, and immunohistochemistry, to detect and quantify the expression levels of β-actin.
Automatically generated - may contain errors
Lab products found in correlation
Anti p gp, by Abcam (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Protein assay dye reagent concentrate, by Bio-Rad (1 mentions)
Chemiluminescence, by Cytiva (1 mentions)
Anti tlr4, by Bioworld Technology (1 mentions)
Anti nlrp3, by Cell Signaling Technology (1 mentions)
Exkine nuclear and cytoplasmic protein extraction kit, by Abbkine (1 mentions)
Fusion solo x, by Vilber (1 mentions)
Anti caspase 1, by Novus Biologicals (1 mentions)
Anti nf κb, by Cell Signaling Technology (1 mentions)
Anti lamin b1, by Cell Signaling Technology (1 mentions)
Anti β tubulin, by Abbkine (1 mentions)
2 protocols using anti β actin
1
Western Blot Analysis of P-glycoprotein
2
Cytosolic and Nuclear Protein Extraction for Western Blotting
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Western blotting was performed as described previously (25 (link)). The cytosolic and nuclear fraction preparation was performed using ExKine™ Nuclear and Cytoplasmic Protein Extraction Kit (Abbkine, China) according to the manufacturer’s instructions. The primary antibodies used were as follows: anti-NLRP3 (1:1,000; Cell Signaling, Danvers, MA, USA), anti-XIAP (1:1,000; Enzo Life Sciences, Farmingdale, NY, USA), anti-ASC (1:1,000; Abbkine), anti-caspase-1 (1:1,000; Novus Biologicals), anti-TLR4 (1:1,000; Bioworld Technology), anti-myeloid differentiation factor88 (MyD88, 1:1,000; Cell Signaling), anti-NF-κB (1:1,000; Cell Signaling), anti-Lamin B1 (1:1,000; Cell Signaling), anti-β-tubulin (1:1,000; Abbkine), and anti-β-actin (1:5,000; Enzo Life Sciences). The membranes were then washed thrice with Tris-buffered saline with Tween-20 (TBST) and incubated with secondary antibodies conjugated with horseradish peroxidase. Immunopositive bands were enhanced with chemiluminescence (Amersham Pharmacia, Piscataway, NJ, USA) and visualized using the Fusion Solo X (Vilber Lourmat, Collegien, France).
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