Cells were lysed by RIPA buffer (100 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, and 1% Triton-X-100) at 4 °C for 10 min, and the cell lysates were harvested by centrifugation at 15,000 rpm for 10 min to obtain the supernatants. The normal brain lysate were purchased from Origene Technologies. Thirty-microgram cell lysates from each group were applied to 10% sodium dodecyl sulfate polyacrylamide gels electrophoresis. Proteins were transferred onto polyvinyldifluoride membranes (Millipore, MA, USA) and blocked with 5% skim milk in TBST for 1 h at room temperature. The antibodies used are anti-DDX3X (GeneTex, San Antonio, TX, USA), anti-estrogen receptor alpha (ERα) antibody (H-184) (cat. No. Sc-7207; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-SP1 antibody (Millipore, Upstate, MA, USA) and β-actin (Santa Cruz Biotechnology, Inc.). Band detection was conducted by enhanced chemi-luminescence and X-ray film (GE Healthcare, Piscataway, NJ, USA).
Enhanced chemi luminescence and x ray film
Manufactured by GE Healthcare
Sourced in United States
Enhanced chemi-luminescence and X-ray film is a laboratory equipment product used for the detection and visualization of proteins, nucleic acids, and other biological molecules. It utilizes chemi-luminescence and X-ray technology to generate images and signals that can be analyzed and quantified.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinyl difluoride membrane, by Merck Group (3 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Anti ddx3x, by GeneTex (1 mentions)
Anti sp1 antibody, by Merck Group (1 mentions)
Mtor antibody, by Abcam (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
P mtor, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
3 protocols using enhanced chemi luminescence and x ray film
1
Western Blot Analysis of Protein Lysates
2
Western Blot Analysis of TELO2, mTOR, and β-Actin
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were lysed by RIPA buffer (100 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, and 1% Triton-X-100) and the cell lysates were harvested by centrifugation at 15,000 rpm for 10 min to remove the debris. The normal brain lysate were purchased from Origene Technologies. Thirty-microgram cell lysates from each group were applied to 10% sodium dodecyl sulfate polyacrylamide gels electrophoresis and proteins were transferred onto polyvinyl difluoride membranes (Millipore, MA, USA). The membrane was blocked with 5% skim milk in TBST for 1 h at room temperature. The antibodies used include anti-TELO2 (Cat. No. ab122722, Abcam, Cambridge, UK), mTOR antibody (cat. No. 2972; Cell Signaling Technology, Beverly, Boston, MA, USA), and β-actin (Santa Cruz Biotechnology, Inc.). Band detection was conducted by enhanced chemi-luminescence and X-ray film (GE Healthcare, Piscataway, NJ, USA).
3
Western Blot Analysis of Cell Signaling
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Cells were lysed by RIPA buffer (100 mM Tris-HCl, 150 mM NaCl, and 0.1% SDS, and 1% Triton-X-100) at 4°C for 10 min, and the cell lysates were harvested by centrifugation at 15,000 rpm for 10 min to obtain the supernatants. Twenty-microgram cell lysates from each group were applied to 8% and 12% sodium dodecyl sulfate polyacrylamide gels electrophoresis. Proteins were transferred onto polyvinyl difluoride membranes (Millipore, MA, USA) and blocked with 5% skim milk in TBST for 1 h at room temperature. The antibodies used are ACTN (Santa Cruz Technology Dallas, TX, USA; SC-17829), p-mTOR (Cell Signaling Technology, Danvers, MA, USA; CST-2971), mTOR (Cell Signaling Technology, Danvers, MA, USA; CST-2972), p-AKT (Cell Signaling Technology, Danvers, MA, USA; CST-9271), AKT (Cell Signaling Technology, Danvers, MA, USA; CST-4691), p-ERK (Cell Signaling Technology, Danvers, MA, USA; CST-4370), ERK (Cell Signaling Technology, Danvers, MA, USA; CST-4695), p62 (Santa Cruz Technology Dallas, TX, USA; SC- 28359), LC3B (Cell Signaling Technology, Danvers, MA, USA; CST-2775), BCL2 (Cell Signaling Technology, Danvers, MA, USA; CST-4223), p-CDC2 (Thr15) (Cell Signaling Technology, Danvers, MA, USA; CST-9111) and CDC2 (Cell Signaling Technology, Danvers, MA, USA; CST-9112). Band detection was conducted by enhanced chemi-luminescence and X-ray film (GE Healthcare, Piscataway, NJ, USA).
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