western-blot

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 ...

1. Run samples out on a gel. For bacterially expressed proteins generally 5 μl is plenty (1ml cell cμlture; cells resuspeded in 50 μl loading buffer). Run the gels (BioRad mini gels) at 195 V ...

1.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 may allow a short circuit and inefficient transfer. Blot t ...

1. 30%储备胶溶液： 丙烯酰胺（Acr） 29.0g　　　　　　　　　　　亚甲双丙烯酰胺（Bis） ...

Storage of lyophilized antibodies All our products are shipped lyophilized (freeze-dried). Unlabeled antibodies are stable without loss of quality at ambient temperatures for several weeks or even a ...

0 128); font-weight: bold;"Western免疫印迹（Western Blot）是将蛋白质转移到膜上，然后利用抗体进行检测。对已知表达蛋白，可用相应抗体作为一抗进行检测，对新基因的表达产物，可通过融合部分的抗体检测。 0 128);"原理与Southern或Northern杂交方法类似，但Western Blot采用的是聚丙烯酰胺凝胶电泳，被检测物是蛋白质，“ ...

Western- or immunoblotting is a commonly employed technique for the detection of protein antigens in complex mixtures. Samples are first separated by SDS-polyacrylamide gel electrophoresis. The separa ...

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

Use alternate buffer system (such as CAPS buffer pH 10.5) with a higher pH. Assemble transfer stack with additional membrane soaked in cathode buffer on the cathode side of gel. This will captur ...

Western Blot详解(原理、分类、试剂、步骤及问题解答)

0 128);"REAGENTSECL 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 ...

Trans-Bot SD Assembly 1. Prepare the transfer buffer. 2. Following electrophoresis equilibrate the gels in transfer buffer. Equilibration facilitates the removal of electrophoresis buffer salts ...

0 128);"Culture bacteria1. Grow glycerol stock A13 (TBP-GST in BL21) in 10mL LB and 10ul ampicillin (1:1000)2. Shake @ 37C ON 0 128);"Induce bacteria3. Adjust culture with LB until ON culture OD is ...

1. Treat cells. For PC12 or NIH 3T3 cells a moderately confluent 60 mm plate is used. 2. Aspirate media. Rinse cells with PBS aspirate. 3. Lyse cells in 200 uL of ...

SJF 4/001) Run gel (in 1:10 running/transfer buffer(10x) and H2O for a total of 1litre) at 150 volts until leading bromophenol blue band is nearing the bottom edge of the glass plates or according to ...

(All steps with agitation unless done O/N at 4oC) Rinse membrane once in TBS-TBlock the membrane with TBS-T/ 5% milk (or HIHS) for 30 min roomtemp (can be left at 4oC O/N here) Rinse membrane briefly ...

Steve Hahn last modified Mon Mar 22 1999Western Blot Probing by ECL1. After electroblotting of protein gel to immobilon membrane (See 0 128);"Novex NuPAGE method) mark the side of the membrane not in ...

0 128);"Buffers: 0 128);"TBS: 25 ml of 1 M Tris-7.5 30 ml of 5 M NaCl bring volume up to 1000 ml with ddwater 0 128);"TBS-T: TBS + 0.5 ml of Tween 20 0 128);"5X SDS-PAGE running buffer:&n ...

Semi-dry Transfer Apparatus Biorad Cat# 170-3940 or equivalent Power Supply 0 - 100 VDC (adj. current to 1 Amp) Immobilon-P transfer membrane 0.45 µm pore size; cut to same size as gel (Millip ...

Wash XCell box blotting module and trays with soap and water before proceeding. 0 128); font-weight: bold;"Reagents:Transfer buffer25 mM Tris HCl190 mM Glycine10% Methanol 0 0); text-decoration: unde ...