蛋白组学

Western Blotting or immunoblotting (Western blotting was dubbed because it is similar to Southern blotting which was invented by and named after the inventor E. M. Southern) allows investigators to de ...

Bacterial Protein Extraction (mini-scale) Using B-Per B-Per (Pierce Cat number: 78248) Extraction of 1.5ml bact.culture OD600nm 1.5-3.5A useful procedure for initial screening of expression conditions ...

Western Blotting or immunoblotting (Western blotting was dubbed because it is similar to Southern blotting which was invented by and named after the inventor E. M. Southern) allows investigators to de ...

WESTERN PROTOCOLA. Preparation of cell lysates Collect cells (confluent T-25) by trypsinization and spin. Lyse the pellet with 100 µl lysis buffer on ice for 10 min. For 500000 cells lyse with 20 ...

Western Blot MaterialsSemi-dry Transfer ApparatusBiorad Cat# 170-3940 or equivalentPower Supply0 - 100 VDC (adj. current to 1 Amp)Immobilon-P transfer membrane0.45 µm pore size; cut to same size as g ...

Cell Lysates For Western Blotting Reagents / Solutions Lysis Buffer:10ml 10% Sodium dodecyl sulphate (SDS)10ml Glycerol10ml b-mercaptoethanol8ml 0.5M Tris pH6.81ml 0.1% bromophenol blue51ml H2O Proto ...

Cell Lysates For Western Blotting Reagents / Solutions Lysis Buffer:10ml 10% Sodium dodecyl sulphate (SDS)10ml Glycerol10ml b-mercaptoethanol8ml 0.5M Tris pH6.81ml 0.1% bromophenol blue51ml H2O Proto ...

Western Blot 1) Run protein gel.Set up blot:2) Prepare two tupperware containers one with dH2O and one with 1/2X Transfer Buffer (TB).3) Remove one glass plate from the gel by twisting a spacer betwee ...

REAGENTS

Western Blotting with Alkaline Phosphatase ConjugatesSAMPLE PREPARATIONFor Protein Concentration Determination of Cell CultureDecant medium from 10cm dish of adherent cells and rinse plate rapidly wit ...

SAMPLE PREPARATIONFor Protein Concentration Determination of Cell CultureDecant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). Aspirate excess PB ...

Western Blotting Using the SemiPhor Semi-Dry Transfer Unit Remove the stacking gel from the gel. Measure the gel and record its size. Do not pre-soak the gel. Do not cut a corner off the gel as this m ...

Studying Protein Interactions by Far-Western BlottingFar-Western blotting was originally developed to screen protein expression libraries with 32P-labeled glutathione S-transferase (GST)-fusion protei ...

Too many bands on a Western blot Western- or immunoblotting is a commonly employed technique for the detection of protein antigens in complex mixtures. Samples are first separated by SDS-polyacrylamid ...

Too many bands on a Western blot Western- or immunoblotting is a commonly employed technique for the detection of protein antigens in complex mixtures. Samples are first separated by SDS-polyacrylamid ...

1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell sample prepared using modified RIPA buffer and transfer the proteins to a nitrocellulose membrane. 2. Wash the nitrocellulose t ...

1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell sample prepared using modified RIPA buffer and transfer the proteins to a nitrocellulose membrane. 2. Wash the nitrocellulose t ...

Troubleshooting Guide: Western BlotProblem: Poor TransferPossible SourceTest or ActionMembrane does not wet uniformlyPre-wet membrane in 100% methanolMW of protein is less than 10000If protein has MW ...

Preparation of Brain Cytosol1. Chill a glass plate on ice. 2. Place brain tissue on the chilled glass plate and cut slices of brain into small pieces. 3. Resuspend the pieces in ice-cold RIPA buffer w ...

Protease Inhibitors from Alberts' Lab Protocols (Kathy Miller/Chris Field)Protease inhibitors are rapidly inactivated in aqueous solutions and should be added to solutions immediately before use. Cock ...