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| Name of Products | HEK293 Cells Stable expressing GPCR protein CXCR4 (Protein accession # NM_003467) |
| Catalogue Number | cAP-1101 |
| Product Format | Frozen Vial |
| Cell Number | > 5x10[5] cells/vial, 2 vials |
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| 1. | HEK293 Cells, stable expressing GPCR protein CXCR4 (HEK293-CXCR4), were developed for screening new human CXCR4 ligands, and antagonists et al based on Aquaporin-Coelenterazine H, chemiluminescent assay. |
| 2. | The cells express AQUAPORIN-GFP in the cytoplasm, G-promiscous protein (Gaqi9) in the cytoplasm, and human CXCR4-GFP on the cell membrane. |
| 3. | Human SDF1, the known ligand for CXCR4, can induce a dose-dependent increase in Coelenterazine H preloaded HEK293-CXCR4 cells. |
| 4. | The whole process for the preparation of the cells in HTP formats takes less than 4 hrs without the hassle to express the target receptor (2-3 days) every time for the assay and the inconsistency of the target expression levels. |
| Cells are offered for Research Use Only (open to be licensed for commercial applications). |
| Frozen Vials in a Dry Ice Package. |
| When you receive the dry ice package with cells in frozen vials, transfer the frozen vials of cells into a -80C freezer for short period storage or a liquid nitrogen tank for long-term storage. |
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| A) | Pre-coating of T25 flasks- Add 4ml each Quick Coating Solution (cAP-01) into a T75 flask to cover the whole surface of the flask, 5 mins later, aspirating the excessive coating solution and the flask is ready to be used. |
| B) | Thaw the frozen cell vial in a 37C water bath first, and then transfer the cells into the pre-coated T75 flask with 15ml of cAP-01B medium, cells usually become confluent with 2-3 days and are ready to be passaged. |
| C) | To passage the cells, rinse the cells in a T75 flask with 10ml HBSS (RT) twice; then add 2ml Trypsin/EDTA (RT) (cAP-23) into one T75 flask; gently aspirating the excessive Trypsin/EDTA solution within 30 seconds. |
| D) | Leave the T75 flask with the cells at RT or 37C for 1 min (most cells usually will detach from the surface within 1-2 minutes, or monitor the cells under a microscope until most of the cells become rounded up, and then gently tap the flask against the bench surface, and the cells will move on the surface of the flask when monitoring under a microscope. |
| E) | Add 10ml Trypsin Neutralization Buffer and spin down the cells with 800g centrifugation for 5 mins. |
| G) | Re-suspend the cell pellet with 30ml cAP-1B medium and transfer 10 ml each into 3 pre-coated T75 flasks (for 1 to 3 subculture ratio). |
| H) | Change medium every 2 or 3days and the cells usually become confluent within 5-7 days (when split at a 1 to3 ratio). |
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| 1. | When cells are nearly confluent, passage the cells as usual, and after spinning down the cells, resuspending the cells with a Phenol Red free medium, containing with 4uM Coelenterazine H. |
| 2. | Counting the cells and add sufficient cells into varial HTP platforms (96-, 384- well plates). After 4 hrs the cells are ready to be used. |
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| Quick Coating Solution | cAP-01 |
| Universal Growth Medium | cAP-01B |
| HBSS w/o Ca2+, Mg2+ | cAP-11 |
| Cell Freezing Solution (FBS) | cAP-22 |
| Cell Freezing Solution (Non-FBS) | cAP-22B |
| Trypsin/EDTA Solution | cAP-23 |
| Trypsin Neutralization Solution | cAP-28 |
| Handling human tissue-derived products is potentially bio-hazardous, despite being tested negative for HIV, HBV and HCV DNA; nonetheless, proper precautions must be taken to avoid inadvertent exposure. |
For customers using our primary cell products, we strongly advise you using our own medium, although similar cell growth medium can be purchased and used for our primary cell products. |
The quality of cells, in term of their growth features stated in our datasheets, will not be guaranteed if NON-ANGIO-PROTEOMIE's Cell Growth Medium is used. |
Due to the sensitive nature of primary cells and cell lines, all quality related issues about the cells products must be reported back to us within ONE month period after receiving the products, no quality warranty (i.e. replacement of cells) will be provided after the ONE-Month period. Thank you for your understanding.
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