Pull-down Target Protein Mass Spectrometry Identification
In the life activities of organisms, the functions of proteins are realized through protein-protein interactions. Therefore, studying new protein interactions can provide new directions for researching the potential functions of proteins. Among many protein interaction identification experiments, Pull-down is commonly used for in vitro protein interaction testing and helps discover new interacting proteins. The Pull-down experiment uses highly purified and enriched bait proteins to capture target proteins that interact weakly or are of low abundance in cells, significantly improving the efficiency of discovering new target proteins. The specific steps of the Pull-down experiment are: express and purify GST, polyHis, or Biotin-labeled bait proteins, use immobilized affinity ligands to stably bind the bait proteins to solid-phase matrices, then mix and incubate the bait proteins with the cell lysate to be tested. During this process, the bait proteins interact and bind with the target proteins. By washing away non-specifically bound contaminant proteins, the target proteins interacting with the bait proteins can be obtained, and then identified through mass spectrometry analysis.
Baite Parker Company uses imported GST pull-down kits to quickly and accurately complete protein interaction experiments. Combined with the company's advanced UPLC-MS mass spectrometry platform, we offer a one-stop technical service for protein analysis based on pull-down, with high sensitivity and accurate resolution. We can either test the protein samples after your pull-down experiment or you can send us the samples, and we will provide a one-stop solution from the pull-down experiment to sample mass spectrometry analysis. You only need to send us your requirements and samples, and we will handle all subsequent matters of the project, including sample pre-processing, pull-down, mass spectrometry analysis, and raw data analysis.
Pull-down Protein Identification Process
Pull-down Protein Identification Process
About Samples
1D SDS PAGE /2D PAGE Gels Samples:
1. Coomassie Blue, SYPRO Ruby, and Silver stain samples can all be used for protein identification experiments.
2. Silver-stained samples may be incompatible with subsequent mass spectrometry analysis. We recommend using the following products or experimental steps for silver staining:
ProteoSilver Plus, Sigma (Product # PROTSIL1 or PROTSIL2)
Dodeca Silver Stain, BioRad (Product # 161-0481 or 161-0480)
3. Do not decolorize silver-stained samples
Liquid Samples (In solution format):
1. If your eluted samples need to be run on a gel, please prepare the samples according to the 1D SDS PAGE sample preparation requirements.
2. If the protein amount in the sample is >10ug, we have no requirements for the sample buffer system.
3. If the protein amount in the sample is <10ug, try to minimize the use of surfactants such as SDS during sample preparation and pay attention to reducing salt concentration.
4. When submitting samples, please also provide the buffer system of the sample and the estimated protein content in the sample.
Sample Transport:
We recommend transporting samples after lyophilization, as proteins in lyophilized samples are very stable.
Liquid samples are recommended to be transported with dry ice; short-distance transport with ice packs is also acceptable.
Note:
When identifying protein samples using mass spectrometry, gloves and hairnets should be worn during sample preparation to avoid contamination of the sample with keratin.
Bilingual Project Report
In the technical report, Baite Parker will provide you with a detailed bilingual (Chinese and English) technical report, which includes:
1. Experimental Steps (Chinese and English)
2. Relevant Mass Spectrometry Parameters (Chinese and English)
3. Mass Spectrometry Images
4. Raw Data
5. Detailed Information on Identified Proteins
One-stop Service for Pull-down Target Protein Mass Spectrometry Identification
You only need to place an order and send samples
Baite Parker One-stop Service Completes: Sample Processing - On-machine Analysis - Data Analysis - Project Report
