Western blot Services

The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used to detect specific proteins in the given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide. The proteins are then transferred to a membrane (typically nitrocellulose or PVDF), where they are probed (detected) using antibodies specific to the target protein.

There are now many reagent companies that specialize in providing antibodies (both monoclonal and polyclonal antibodies) against tens of thousands of different proteins.[3] Commercial antibodies can be expensive, although the unbound antibody can be reused between experiments. This method is used in the fields of molecular biology, biochemistry, immunogenetics and other molecular biology disciplines.

We can provide WB service. The most steps is followed.
1 Sample preparation
  Tissue or cell culture is accepted.
2. Assay the concentration of the sample protein by BCA method.
3. Gel electrophoresis (SDS-PAGE).
4. Transfer

Proteins are moved from within the gel onto a membrane made of nitrocellulose by using a Bio-Rad Trans-Blot under 30mA and 4°C overnight.
5. Blocking
  Placethe membrane in 5% non-fat dry milk buffer for 1 hour at least to prevent non-specific binding.
  After blocking, a dilute solution of primary antibody (generally between 0.5 and 5 micrograms/mL) is incubated with the membrane under room temperature for 1.5 hour.
  Expose the membrane to secondary antibody conjugated HRP. Incubate under room temperature for 1.5 hour. ( GAPDH-conjugated HRP is added to assay the GAPDH concentration at the same time.)
6. Chemiluminescent detection
7. Data analysis
  Correction errors by using GAPDF as the internal control.
8. Report with detailed experimental methods and WB experimental result with related datas.

Price
1500RMB / membrane (exclude the primary antibody)
（8 samples and internal control on one membrane）
Time: 2-3 weeks

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