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CypK

CAS No. : 1610703-09-7

MCE 站：CypK

产品活性：CypK (N-Cyclopropene-L-Lysine) 是赖氨酸的环丙烯衍生物，通过基因编码扩增有效地整合到抗体中。CypK 是一个*小的生物正交柄，用于创建稳定的治疗蛋白偶联物。

研究领域：Others

作用靶点：Others

In Vitro: Guidelines (Following is our recommended protocol. This protocol only provides a guideline, ａnd should be modified according to your specific needs).
CypK assay (express the antibody):
1. Thaw a vial of HEK suspension cells in a 250 mL flask containing 50 mL of expression medium supplemented with 100 units/mL penicillin, 100 g/mL streptomycin, ａnd 250 ng/mL amphotericin B. Keep the cells at 37 ^oC with 8% CO₂ in humidified incubators equipped with a shaker at 125 rpm. Split cells to 0.3-0.5 x 10 cells/mL (every 2-3 days) at least 2 times before transfecting.
2. When a density of 2.5 x 10 cells/mL is reached (2-3 days after splitting), prepare a fresh solution of 100 mM CypK. For this purpose, weigh 64 mg of CypK, add 2.5 mL of 0.1 sodium hydroxide, vortex, spin down to recover all undissolved particles ａnd sonicate.
3. Add 2.5 mL of CypK (100 mM in 0.1 M NaOH) to 42.5 mL of expression medium supplemented with antibiotics. Mix well, add 250 L of 0.1 M HCl, ａnd sterilize using a 0.22 m filter.
4.Dilute 50 g of HC ａnd LC pKym1 plasmids to 2.5 mL with reduced serum medium. In a separate tube, dilute 135 L of transfection reagent to 2.5 mL with reduced serum medium.
5. Five minutes after preparing the solutions, mix the plasmids ａnd the transfection reagent solution ａnd incubate for 20 min to allow the formation of complexes between the DNA ａnd the transfection reagent.
6. In the meantime, centrifuge 125 million cells at the target density for 5 min at 500 x g, resuspend with the expression medium containing CypK ａnd add the DNAtransfection reagent mixture.
7. After incubating cells for 20 h, add 250 L of transfection reagent enhancers included in the kit.
8. Harvest antibodies from the supernatant 6-7 days after addition of CypK (no change of medium is required during expression).

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