摘要:SAMPLE PREPARATIONFor Protein Concentration Determination of Cell Culture1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). 2.Aspirate exces[阅读全文:]
摘要:SAMPLE PREPARATIONFor Protein Concentration Determination of Cell Culture1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). 2.Aspirate exces[阅读全文:]
摘要:REAGENTS ECL Western blotting kit (Amersham Life Science; cat# RPN2108): contains second antibodies for both mouse and rabbit, substrate and milk blocker (the milk blocker is not normally used when us[阅读全文:]
摘要:Western Blot 1、Run protein gel.Set up blot:2、Prepare two tupperware containers, one with dH2Oand one with 1/2X Transfer Buffer (TB).3、Remove one glass plate from the gel by twisting a spacer between t[阅读全文:]
摘要:Materials Blot Cell BA 83 0.2 µm pore nitrocellulose sheets Buffer, PBS-Tween 20 Antigenic proteins, antibodies, and horseradish peroxidase labeled antiglobulins Procedure Run an electrophoretic[阅读全文:]
摘要:Preparation of Brain Membrane Fractions for Western Blot AnalysisAll the procedures should be done at 4ºC using precooled reagents. For rat samples, immediately remove brain from the cranium into[阅读全文:]
摘要:Reagents / Solutions Lysis Buffer:10ml 10% Sodium dodecyl sulphate (SDS)10ml Glycerol10ml b-mercaptoethanol8ml 0.5M Tris pH6.81ml 0.1% bromophenol blue51ml H2OProtocolSpin down 106 cells and wash, if [阅读全文:]
摘要:The following tips can be used to overcome the most common problems encountered during Western blotting. Smeared Pattern or Distorted Bands Uneven contact between gel and membrane: cassettes used shou[阅读全文:]
摘要:TGEK Base50 mM Tris10% vol Glycerol1 mM EDTA100 mM KCl(add for std lysis buffer):PMSFBenzamidineLeupeptinAprotinin0.5% NP40(add for solubilization buffer)1% NP400.1% Triton X1000.1% SDS0.5% NaDeoxycho[阅读全文:]
摘要:SolutionsElution Buffer50 mM Tris 7.5 5 ml 1M Tris pH 7.50.1% SDS 0.1 g SDS0.1 mg/ml BSA 1.0 mg BSA0.25 mM EDTA 50 ml 0.5M EDTA2.5% glycerol 2.5 ml glycerolup to 100 ml with QFor every 10 ml add:10 ml[阅读全文:]