Immunoglobulin igg

HAuCl₄·3H₂O, K₂CO₃, Immunoglobulin (IgG), PVP-40, BSA, NaCl, Tween-20, EDTA, NaN₃, Sucrose, Tris, β-cyclodextrin, and staphylococcal protein A (SPA) were purchased from Sigma (St. Louis, MO, USA). A lipopolysaccharide extraction and purification kit was purchased from Intron Biotechnology Company (Chengnan, Korea). The water used in this study was deionized. The cover, detection box, and nitrocellulose filter membrane (NC) were all purchased from Advanced Microdevices (Haryana, India).

HEK-293T cells were cross-linked with 1% formaldehyde for 10 min at room temperature, washed with ice-cold phosphate-buffered saline (PBS), and stop-fix solution containing glycine (125 mM), aprotinin (1 μg/ml), pepstatin A (1 μg/ml) and Pefabloc (1 mg/ml) was added to neutralize the DNA-protein cross-linking for 5 min at room temperature. The samples were pelleted by centrifugation at 2,500 × g and 4°C for 2 min and washed with ice-cold PBS. Chromatin was immunoprecipitated with immunoglobulin IgG (Sigma) and anti-EGR1 (catalog no. 4154S; CST). The association of EGR1 with ADAM10 was measured by qRT-PCR with the following primers: forward, 5′-CAGGCCTAGCAGCACGGGA-3′, and reverse, 5′-TCCCTTGCTCGTTCCCTCT-3′.