蛋白质技术

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 ...

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)