Question: What are the differences between the tube-based and plate-based CMO labeling protocols for using the 3' CellPlex kit for Cell Multiplexing?
Answer: The tube-based CMO labeling protocol (CG000391) is recommended for CellPlex experiments involving 12 or fewer samples. The plate-based CMO labeling protocol (CG000426) is suitable for high throughput plate-based assays with large sample numbers, such as large-scale drug screens, but can be also used for low input samples (see updated plate-based DP).
Prior experience with high throughput plate-based assays is recommended for success with the plate-based CMO labeling protocol.
The minimum sample input requirements differ between protocols. At least 100,000 cells or nuclei from fresh cells/tissues per sample are required for the tube-based CMO labeling protocol. In contrast, at least 10,000 cells per sample are required for the plate-based CMO labeling protocol.
Additional differences between the protocols are outlined in this table:
Please note that each protocol should be followed as it is written. We do not recommend interchanging aspects of the protocols:
- Using lower cell inputs with the tube-based protocol may lead to insufficient cell recovery & difficulty removing the supernatant during each wash step (pellet may not be visible)
- Using higher cell inputs with the plate-based protocol may lead to insufficient washing (the lower wash volumes in this protocol may be insufficient for higher cell loads)
- A vacuum pump is required for plate-based protocol, for efficient removal of supernatant
Products: Single Cell Gene Expression, CellPlex