Ramos (HiBiT) TCK Bioassay Technical Manual
Instructions for Use of Product(s)
JA1411, JA1415, JA2461, JA2465
Literature # TM749
HiBiT TCK Bioassays measure the death of target cells engineered to express a HiBiT fusion protein. HiBiT is an 11 amino acid peptide that binds with high affinity to LgBiT to form NanoBiT® luciferase. When dead or dying cells release the HiBiT fusion protein into the extracellular medium, it binds to the cell impermeable LgBiT, which is provided with the furimazine substrate in a nonlytic, homogeneous detection reagent. In coculture experiments, the luminescent signal will be proportional to target cell death alone, with no signal contribution arising from the death of effector cells.
Ramos (HiBiT) TCK Bioassay is provided in a thaw-and-use format, as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell culture and propagation.
When Ramos (HT-HiBiT) target cells are cocultured with appropriate effector cells (e.g., NK cells or cytotoxic T cells) they can be used to specifically monitor the activity of biologics directed against TAA expressed on the Ramos Cells (HT-HiBiT), such as CD19, CD20 or CD22. When Ramos Cells (HT-HiBiT) are used in assays with biologics against TAA they do not express, the target cells are not lysed and no increase in luminescent signal is observed.
This Technical Manual is relevant for Cat.# JA1411 and JA1415; JA2461 and JA2465 (Korea).
Summary of Changes
The following changes were made to the 3/25 revision of this document:
1. In Section 2, quantities of RPMI 1640 Medium and FBS in the kits changed.
2. In Section 3, text about the product label was revised.
3. In Section 5.A, the amount of CD8+ T cell TDCC assay buffer to prepare was reduced.
4. In Section 5.D, Steps 6–10 were revised and are now Steps 6–8.
5. In Section 6.D, Steps 6–11 were revised and are now Steps 6–8.
6. Some subheadings were updated to include the day on which that protocol is performed.
7. Figures 9 and 12 were revised.
8. Patent statements were removed.
Revised 3/25.