- Primary Structure Analysis
- High-resolution mass spectrometry molecular weight
- MALDI TOF mass spectrometry analysis
- N-terminal sequence analysis
- C-terminal sequence analysis
- N/C terminal sequence analysis
- Analysis of the K deletion ratio at the C-terminus of antibodies
- LC-MS/MS protein full sequence validation
- Peptide coverage / Peptide spectrum analysis
- Protein peptide profile determination
- Amino Acid Composition Analysis
- Extinction coefficient analysis
- View more
- Advanced Structural Analysis
- Charge Heterogeneity Analysis
- Impurity Analysis
- Native Mass Spectrometry
- SDS-PAGE protein purity analysis
- Protein purity analysis (size exclusion/reverse phase chromatography)
- Host Cell Protein Residue (HCP) Analysis Service
- Antibody-Drug Conjugates (ADCs) Analysis
- Protein content analysis
- Product-related impurity analysis
- Analysis of other process-related impurities
- View more
- Special Analysis
- Primary Structure Analysis
What preparations do I need for Western Blot (WB) experiments?
Before conducting a Western Blot (WB) experiment, several key preparatory steps need to be completed:
1. Experimental Design:
- Identify the target protein and suitable antibodies (primary and secondary antibodies).
- Select an appropriate loading control (e.g., GAPDH, β-actin, etc.).
- Determine the sample type (cell lysate, tissue extract, etc.).
2. Sample Preparation:
- Collect and prepare samples (cell culture, tissue samples, etc.).
- Lyse cells or tissues using an appropriate lysis buffer to extract proteins.
- Determine protein concentration (e.g., Bradford, BCA protein assay).
3. Electrophoresis Preparation:
- Prepare SDS-PAGE gel (can be pre-cast or homemade).
- Prepare electrophoresis buffer.
- Prepare samples (add sample buffer and boil).
4. Transfer Preparation:
- Prepare PVDF or nitrocellulose membrane.
- Prepare transfer buffer.
- Prepare transfer equipment (such as semi-dry or wet transfer system).
5. Membrane Blocking and Antibody Incubation:
- Prepare blocking buffer (such as 5% non-fat milk or BSA solution).
- Prepare primary and secondary antibody solutions.
6. Detection:
- Prepare a detection system suitable for the used secondary antibody (chemiluminescence, fluorescence, etc.).
7. Other Materials and Equipment:
- Microcentrifuge tubes, pipettes, heat block or water bath, electrophoresis tank, power supply, membrane holders, shaker, etc.
- General laboratory consumables (gloves, paper towels, washing liquids, etc.).
8. Experimental Record and Data Analysis:
- Prepare a lab notebook to meticulously record each step of the experiment.
- Determine methods and software for data analysis.
Before performing the WB experiment, it is recommended to carefully read the relevant operation manuals. If necessary, a small-scale pre-experiment can be conducted to optimize conditions. Preparing troubleshooting strategies for potential issues during the experiment is also very helpful.
Bio-Techne--A leading service provider for bioproduct characterization and multi-omics mass spectrometry analysis.
Related Services:
Protein Interaction Mass Spectrometry Analysis
Co-immunoprecipitation (CO-IP) combined with mass spectrometry for protein interaction analysis
SILAC combined with co-immunoprecipitation and mass spectrometry for protein interaction analysis
Crosslinking Protein Interaction Analysis
How to order?
