1.How much tissues or cells should I provide for proteomic analysis?
The amount of tissue or cells required for proteomic analysis may vary depending on the type of sample and the abundance of the protein of interest. As a general guideline, the following minimum amounts of starting material are recommended:
1) For animal tissue, at least 15 mg should be provided.
2) For plant tissue, at least 100 mg should be provided.
3) For cells, a minimum of 1.5 x 106 cells and 7 μL of routine cell precipitate should be provided as a reference.
2.How do I collect and send cell samples?
1) Digest the adherent cells with trypsin or scrape them off with a cell scraper and collect them into 1.5 mL centrifuge tubes.
2) Wash the cells twice with pre-cooled PBS and collect them by centrifugation at low speed (600 g) at 4℃.
3) Rapidly freeze the resulting cell precipitates using liquid nitrogen and store them in a -80℃ freezer until shipment.
4) When shipping the cell samples, pack them with dry ice to ensure they remain frozen during transportation.
It's important to note that shipping biological samples may be subject to various regulations and requirements, depending on the destination and mode of transport. Therefore, it's advisable to consult with the shipping company or relevant authorities to ensure compliance with all applicable regulations.
3.How do I collect and send animal samples?
When collecting clinical or animal tissue samples, it is important to clean them with pre-cooled PBS after accurate sampling until there is no obvious red color in the PBS solution. For tissue over 100 mg, it should be cut and cleaned. Next, the samples should be rapidly frozen in liquid nitrogen and stored in a -80℃ refrigerator. Finally, the samples should be sent with dry ice.
4.How much clinical FFPE or paraffin slices should I provide for proteomic analysis?
For proteomic analysis using clinical FFPE or paraffin slices, it is recommended to provide slices with a thickness of 5-10 μm for FFPE samples. For paraffin slices, there is no specific specification, but it is recommended to provide at least 4 slices with a size of 4cm2 and a thickness of 10 μm or greater. The overall volume of the provided samples should be greater than 15 mm3.
5.How much puncture samples should I provide for Proteomic Analysis?
The amount of puncture samples required for proteomic analysis can vary depending on the specific experiment and sample type. Common puncture needle specifications range from 6G to 20G, with an inner diameter between 0.6-2mm. The length of the needle depends on the sample type, for example, bone marrow biopsy samples may require a length greater than 1.5cm, while common tissue puncture samples may be slightly shorter. After obtaining the tissue, it is recommended to clean it with pre-cooled PBS until there is no obvious red color in the PBS solution. Then, the tissue should be rapidly frozen in liquid nitrogen and stored at -80℃ until shipment. The amount of tissue required can vary, so it is best to consult with the testing facility or experiment protocol to determine the optimal amount of sample needed.
6.How much tissue or cells should I provide for Lactylome Analysis?
The amount of protein required for PTM quantitative analysis is higher than that of proteomic analysis. It is recommended that you provide as many samples as possible. The amount of tissue or cells needed can vary depending on the type of sample and protein abundance. However, the minimum amount of sample required is:
1) Animal tissue: not less than 80 mg;
2) Plant tissue: not less than 500 mg;
3) Cells: The minimum amount of routine cell precipitates is 70 μL, and the number of cells should be at least 2x107 as a reference.
7.Can Serum, Blood Cells, B Cells, T Cells, etc. be Used for Proteomic Research?
Yes, erythrocytes, leukocytes, lymphocytes (containing B cells, T cells, etc.), and serum (only human serum can remove high abundance proteins) can be used for proteomic research.
8.Protein Extraction of the Sample (Foreign Customers Need to Extract Protein Themselves)
For infectious or pathogenic bacteria such as Mycobacterium tuberculosis or viruses of infected samples (refer to the classification of pathogens/viruses for details), customers are responsible for extracting the protein solution. For other samples, we recommend that you send them directly to us on dry ice, and we will be responsible for protein extraction. Protein extraction is the most basic step in the whole proteomic analysis and is crucial as it directly affects the results of subsequent experiments.
9.Protein Extraction Method for Pathogenic Bacteria and Virus-Infected Cells or Tissues
Protein Extraction Method for Pathogenic Bacteria and Virus-Infected Cells or Tissues
This method is suitable for Class II and III pathogenic bacteria and tissues infected with these two types of bacteria. For Class I pathogenic bacteria samples, technical services are not available. Class IV samples are not required at this time. For specific classification standards, please refer to Jingjie Biological Pathogen Microorganism List.
For cell, tissue, bacterial, and fungal samples:
1) Remove the samples from -80 °C and add lysis solution (containing 1% SDS) at a volume equal to twice the sample volume (or adjust according to the actual situation).
2) Sterilize the samples at 121 °C for 30 min using moist heat (or heat at 100 °C for 30 min with the lid open).
3) Grind the samples into powder by adding liquid nitrogen to the lysate (cellular samples do not need to be manipulated).
4) Sonicate the samples in ice water for 3 min using the following parameters: sonication for 3 sec, stop for 5 sec, power at 220W.
5) Place the samples in a water bath at 100 °C for 20 min for thermal lysis.
6) Seal the samples with sealing film, place them in self-sealing bags or 50 ml centrifuge tubes, and send them directly to our company in a dry ice environment (do not centrifuge, retain the residue).
For prion samples:
1) Autoclave the samples at 134 °C for 60 min to inactivate them.
2) Grind the samples in liquid nitrogen and add an appropriate amount of lysis solution (containing 1% SDS) to the ground powder (except cell samples).
