![]() ![]() Replace with a transfer buffer (25 mmol/L Tris, 192 mmol/L Glycine, 10% MeOH) that does not contain EDTA, agitate for 10 minutes, and transfer onto PVDF membrane or nitrocellulose membrane. Immerse the gel in a transfer buffer (25 mmol/L Tris, 192 mmol/L Glycine, 10% MeOH) containing 10 mmol/L EDTA, and agitate gently for 10 minutes. Yes: However, zinc in the gel has to be removed by EDTA treatment to achieve optimal transfer efficiency to the membrane. Can the product be used for Western blot? Be careful with handling as the container will be hot. Replace deionized water and repeat the same procedure 2-3 times. Transfer the stained gel in 100 mL of deionized water in the container, add a few rolls of Kim-Wipes, and heat it in a microwave for a few minutes. CBB-stained gel cannot be de-stained clearly.Ī microwave can be used when de-staining to give you a clearer gel. What gel staining methods can we use?ĬBB, negative, silver, and florescent staining methods can be used. is suitable for Invitrogen ® XCell SureLock ® Mini-Cell Tank.is suitable for Bio-Rad Mini-PROTEAN Tank.Phos-tag Precast Gels are available in three cassette sizes,, and. What electrophoresis tanks are the Phos-tagTM Precast Gels compatible with? The level of protein X phosphorylation changed over time after X-ray irradiation. Protein accumulation for p53 was at the highest level 4 hours after X-ray irradiation. Detection was performed by chemoluminecence. The membrane was blocked with 2% Milk/TBS-T, and was reacted with primary antibodies (upper image: p53, lower image: cell-cycle proteins). Gentle agitation of the gel was performed in a transfer buffer containing 10 mmol/L EDTA, and proteins were transferred to PVDF membrane. Cell extracts were prepared, and SDS-PAGE was performed in 13 wells using 50 μmol/L of 10% SuperSep™ Phos-tag™ X-ray irradiation (5 Gy) was performed for human lung cancer cell line Lu99, and cells were collected across different time points. of Radiation Biology, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, University of Tokyo Procedure Atsushi Enomoto, Molecular Radiology / Sec. Time-dependent Change in the level of phosphorylation Data was provided by courtesy of Dr. In addition, the results demonstrated the level of dephosphorylation across different time points. The results confirmed separation of phosphorylated and dephosphorylated β-casein. Dephosphorylation of β-casein was performed over different time periods. ![]()
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