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Purification of TFIIA from E.coli Linda WarfieldHahn Lab2003 Volumes given are for 2L of cells for each subunit (Toa1 and Toa2). Toa1 and Toa2 subunits are expressed separately in cultures of OD0.6 fo ...

To concentrate proteins for analysis by SDS PAGE: If a small amount of protein is to be precipitated (less than a few micrograms)add Insulin as a carrier protein (10 micrograms of Sigma insulinI-5500p ...

Summary WCE Extracts are prepared from strain SHY282 (TFC4-Flag-EE (HpaI)).TFIIIC complex is first purified by flag affinity: TFC4 contains a single copy of the flag epitope.The flag purified TFIIIC i ...

Horvath and RiezmanYeast1994; Gottschling Lab Sample Buffer: 10 ml 0.06M Tris-HClpH 6.8 0.6 ml 1M Tris 6.8 10% (v/v)glycerol 2 ml 50% glycerol 2% (w/v)SDS 2 ml 10% SDS 5% (v/v)2-mercaptoethanol 0. ...

Adapted from YaffeM.P.and SchatzG.PNAS1984814819-4823. 1.Grow 25mls yeast cells to 5x10E6. 2.Sequentially spin down cells in a 15 ml polypro tube. 3.Wash cells with 1ml ice cold ddH2Otransferrring to ...

Solutions Elution Buffer 50 mM Tris 7.5 5 ml 1M Tris pH 7.5 0.1% SDS 0.1 g SDS 0.1 mg/ml BSA 1.0 mg BSA 0.25 mM EDTA 50 ml 0.5M EDTA 2.5% glycerol 2.5 ml glycerol up to 100 ml with Q For every 10 ml a ...

TGEK Base 50 mM Tris 10% vol Glycerol 1 mM EDTA 100 mM KCl (add for std lysis buffer): PMSF Benzamidine Leupeptin Aprotinin 0.5% NP40 (add for solubilization buffer) 1% NP40 0.1% Triton X100 0.1% SDS ...

高性能的实时定量PCR和RT-PCR要求高性能的系统。易于使用的qPCR 超级混合物、一步和两步定量RT-PCR系统、以及荧光检测平台，在一个较宽的动态范围内提供更高的灵敏度和特异性，它们利用了最先进的技术，包括： 使用抗体介导热启动技术的Platinum ®　Taq DNA Polymerase提供在室温下组合PCR反应时更高的特异性和产量。 增加了热稳定性和半衰期的SuperScri ...

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 sh ...

Preparation of Brain Membrane Fractions for Western Blot Analysis All the procedures should be done at 4ºC using precooled reagents. For rat samples immediately remove brain from the cranium into ...

Reagents / Solutions Lysis Buffer: 10ml 10% Sodium dodecyl sulphate (SDS) 10ml Glycerol 10ml b-mercaptoethanol 8ml 0.5M Tris pH6.8 1ml 0.1% bromophenol blue 51ml H2O Protocol Spin down 106 cells and ...

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 electrophore ...

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 betwee ...

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 ...

Western Blotting with Alkaline Phosphatase Conjugates SAMPLE PREPARATION For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidl ...

Western Blotting with Biotinylated Antibodies SAMPLE PREPARATION For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with p ...

SAMPLE PREPARATION For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). 2.Aspirate ex ...

Sample Preparation For Protein Concentration Determination of Cell Culture 1.Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). 2.Aspirate e ...

Running Protein Gels Solutions 10X Running Buffer (0.25 M Tris 1.92 M glycine 1% SDS) 121 g Tris 577 g glycine 40 g SDS ddh20 to 4 L (check pH at 1:10 dilution (pH=~8.8)). 5X Sample Buffer (0.3125 M T ...

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