DNA pull down-MS is a powerful tool to study DNA-protein interactions in vitro. Biotin-labeled DNA fragments are bound to streptavidin magnetic beads and then incubated with cellular nuclear proteins. The protein product is then purified and eluted to detect whether the specific protein binds to the target DNA fragment by western-blot; mass spectrometry is used to identify the proteins that may interact with the DNA fragment (e.g., a specific transcription factor/histone, etc.).
The interaction between biotin and streptavidin is the most intense non-covalent interaction known. Among them, activated biotin can be coupled to almost all known biological macromolecules mediated by protein cross-linking agents. Therefore, after labeling nucleic acids with biotin, a variety of biomolecular complexes can be purified using the affinity interaction between biotin and streptavidin.
DNA pull down experiments begin with the design of a DNA probe for the target region, which is labeled with dethiobiotin and bound to streptavidin coupled to a magnetic bead. Nucleus extracts are incubated with the beads-DNA probe to purify the protein complex bound to the target DNA fragment, which is then eluted to obtain the bound protein of the DNA fragment.
Screening of interacting proteins (such as transcription factors, etc.) of target DNA fragments.
Relying on a complete and advanced proteomics platform, Creative Proteomics has rich project experience in mass spectrometry identification of trace proteins such as DNA pull down products, and has unique insights into subsequent bioinformatics analysis. Creative Proteomics can provide comprehensive guidance around the overall topic of DNA-binding proteins, including upstream and downstream experimental design and multi-method validation of binding proteins.
|Project||Customer Provided||Result Delivery|
|DNA pull down||1. Gene information (species, sequence)
2. cell line and culture information
3. Protein information to be validated
4. Protein antibodies
|1. Raw results figure(including Input, positive and negative control results)
2. Complete experimental report (including experimental materials, methods, and analysis of results)
|DNA pull down MS||1. Gene information (species, sequence or ID)
2. Cell line and culture information
|1. Raw results figure (SDS-PAGE graph of input and eluate)
2. Mass spectrometry search results
3. complete experimental report (including experimental materials, methods, and analysis of results)
Probes designed for a wide range of lengths
Protein extracted from fresh tissues and cells, maintaining natural conformation
High specificity and low false positive rate
Suitable for eukaryotes and prokaryotes
Short experimental cycle time, high throughput, high sensitivity
Live cells: > 108.
Fresh tissue: > 0.5g.
Creative Proteomics has independent laboratories for proteomics, interactomics, and molecular cells. The laboratories are equipped with large precision instruments such as liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), isoelectric focusing instrument and various specialized analytical testing equipment. We realize the traceability of data results and ensure the accuracy of experimental results.
Creative Proteomics provides high quality technical services to biopharmaceutical companies, universities, research institutions, hospitals and individuals. We have assisted our clients in publishing papers in high level journals such as Nature methods, Journal of Hematology & Oncology, etc.
Thank you for choosing our Pull Down service. If you have any related questions, please contact our staff and we will answer and serve you as soon as possible.