Organoids were dissociated by a homogenizer and washed with 1× PBS. Homogenized organoids were extracted in lysis buffer (1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 150 mmol/L NaCl in 50 mmol/L Tris [pH 8.0], Sigma-Aldrich; and 1× proteinase inhibitor mixture, Roche). The extracted protein was separated by 12% SDS-PAGE and transferred to nitrocellulose membranes. The membrane was probed with the following primary antibodies: NURR1 (1:200, Santa Cruz, sc-991), VMAT2 (1:1,000, Abcam, AB1598P), PITX3 (1:1,000, Invitrogen, 382850), pS129-α-synuclein (1:500, Abcam, AB9850), cleaved caspase-3 (1:500, Cell Signaling, 9661s), TXNIP (1:500, Thermo, 40-3700), Phospho-ERK1/2 (1:1,000, Cell Signaling, 4370s), ERK1/2 (1:1,000, Cell Signaling, 4695s), Phospho-p38 (1:500, Thermo, MA5-15177), p38 (1:500, Antibodies online, abin2957701), and β-actin (1:1,000, AbFrontier, LF-PA0207). Representative images are shown of western blots performed using Chemidoc TRS+ with Image Lab software (Bio-Rad).
Chemidoc trs
Manufactured by Bio-Rad
The Chemidoc TRS+ is a gel documentation system designed for imaging and analyzing gel-based protein and nucleic acid samples. It provides high-quality image capture and analysis capabilities, enabling researchers to visualize and quantify their experimental results.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by AbFrontier (2 mentions)
Ps129 α synuclein, by Abcam (1 mentions)
Phospho p38, by Thermo Fisher Scientific (1 mentions)
Txnip, by Thermo Fisher Scientific (1 mentions)
Phospho erk1 2, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Proteinase inhibitor mixture, by Roche (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Apoe4, by Merck Group (1 mentions)
Igfbp 3, by Santa Cruz Biotechnology (1 mentions)
Anti aβ 6e10, by BioLegend (1 mentions)
2 protocols using chemidoc trs
1
Molecular Analysis of Dopaminergic Organoids
2
Alzheimer's Disease Biomarker Analysis
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Samples of AD-patient iNs were washed with 1 × PBS and then lysed in RIPA buffer containing 1% NP-40, 0.5% DOC, 0.1% SDS, and 150 mmol/l NaCl in 50 mmol/l Tris (pH 8.0) supplemented with 1 × proteinase inhibitor mixture (GenDepot, Barker, TX). Following the previously published protocol [21 (link)], the supernatant for Aβ analysis was electrophoresed on 12% sodium dodecyl sulfate–polyacrylamide gel and transferred to nitrocellulose membranes (GE Healthcare Bio-Sciences, Piscataway, NJ). Primary antibodies (anti-Aβ (6E10), 1:300, Biolegend; apoE4, 1:1000, Millipore; IGFBP3, 1:1000, Santa Cruz, Dallas, TX; β-actin, 1:1000, AbFrontier, Seoul, Korea) were applied overnight at 4 °C. Representative images were obtained using Chemidoc TRS + with Image Lab software (Bio-Rad Laboratories, Hercules, CA).
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