Technical advantages
Protein expression
Prokaryotic expression system
Step 1. E.coli ex
The ex
At least 5 positive clones were selected and amplified in LB medium to induce the ex
SDS-PAGE electrophoresis was used to detect the ex
Providing ex
Supply:ex
Delivery time: one week
Step 2. Target protein ex
1L recombinant bacteria were cultured in shake flask to induce the ex
The recombinant protein was purified by various chromatography methods such as affinity ion exchange, hydrophobic and gel filtration.
Remove unwanted purification tags using proteases(Tag), The desired target protein was obtained by re-purification (Optionally, appropriate protease cutting sites should be added when constructing ex
Testing the results of each step though SDS-PAGE electrophoresisand and controls the quality of the final product;Verify the correctness of target protein though Western-blot.
Supply:Purified good target protein 5~10mg and purified report. The final protein which contains purification Tag (Tag) or excision purification accounding to your requirement, >95% purity.
Delivery time: 2~3weeks
Note:
1. There is no charge if the target protein is failed, Both parties negotiate the fees(20%~50% discount) if the protein don't meet your requirement but you stil accept it.
2. There is an additional cost if you need the detailed purification process or conditions and results of each step.
3. We supply 1-3mg target protein basis on the ex
4. If the delivery form is protein liquid, the default storage buffer is Tris/PBS-ba
So we can't guarantee the biological activity of the final product, However, we are committed to preparing the target protein in a way that maximizes its activity, We can provide approximately 50μg purified samples for customers to test activity if target protein activity must be required. We no longer supply the target protein to you and only charge 30% of the cost of this step if the sample is unqualified.
5. It is up to seven samples when Western-blot testing because positive control, negative control and Marker will be added.
This step includes only one free test, Additional charges apply if more tests are required. We can provide you with anti-HIS-TAG primary antibodies free of charge, We don't guarantee Western-blot because false positive or false negative may occur. But we can redo it one time for free if it doesn't work out.
Step 3. Reprocessing and detailed detection of target proteins
The purified protein products with high biological activity requirements were investigated by renaturation, endotoxin removal, sterilization, freeze-drying and other further processing, and the quality of the target protein was tested in detail by HPLC, mass spectrometry and other methods, and the target gene was subcloned into a suitable ex
Service content:
1. Renaturation research: Using up to 18 different renaturation buffer systems, small renaturation tests are conducted on target proteins to provide renaturation samples for customer testing. According to customer requirements, a small amount of reduced protein can be prepared according to the reduced condition of one of the samples, and specific reduced buffer formula and reduced process can be provided.
2. Endotoxin removal
3. Sterilization by filtration
4. lyophilization
5. HPLC test
6. Mass spectrometry test
Delivery time:1~2weeks
Step 4. Large-scale preparation
Provide customer: qualified target protein and service report.
