ADAMII CD34 has dilution protocol for different sample types based on the number of white blood cells (WBCs) from complete blood count (CBC)
Please refer to the following standard to dilute.
Table. Dilution protocol summary
Sample type |
WBC standard |
Dilution procedures |
Dilution reagents |
Dilution factor |
| HPC-A | <150,000 cells/ul | before staining | PBS | see case 1, 2 |
| MPB | <35,000 cells/ul | before loading | 1x RBC lysis Buffer | see case 3, 4 |
| Cord Blood | No need | - | - |
Example of dilution protocol
Case 1. HPC-A. WBC 200,00 cells/ul
Your HPC-A sample has 200,000 cells/ul of WBCs according to CBC count.
--> You have to dilute it by 1/2 to fall within the standard of <150,000 cells/ul
Aliquot 100~ 500 cells/ul (recommend 500).
Then, dilute it by 50% with PBS.
Proceed with staining for 20 minutes and lysis for 10 minutes. Then load the sample.
Before counting, set the dilution factor to 2 as the sample is diluted by 1/2.
Case 2. HPC-A. WBC 330,00 cells/ul
Your HPC-A sample has 330,000 cells/ul of WBCs according to CBC count.
--> You have to dilute it by 1/3 to fall within the standard of <150,000 cells/ul
Aliquot 100~ 500 cells/ul (recommend 500).
Then, dilute it by 1/2 with PBS.
Proceed with staining for 20 minutes and lysis for 10 minutes. Then load the sample.
Before counting, set the dilution factor to 3 as the sample is diluted by 1/3.
Case 3. MPB WBC 55,000 cells/ul
Your HPC-A sample has 55,000 cells/ul of WBCs according to CBC count.
--> You have to dilute it by 1/2 to fall within the standard of <350,000 cells/ul
Aliquot 100~ 500 cells/ul (recommend 500).
Proceed with staining for 20 minutes and lysis for 10 minutes.
Dilute it by 1/2 with 1x RBC lysis buffer.
Then load the sample.
Before counting, set the dilution factor to 2 as the sample is diluted by 1/2.
Case 4. MPB WBC 81,000 cells/ul
Your HPC-A sample has 81,000 cells/ul of WBCs according to CBC count.
--> You have to dilute it by 1/3 to fall within the standard of <35,000 cells/ul
Aliquot 100~ 500 cells/ul (recommend 500).
Then, dilute it by 1/3 with PBS.
Proceed with staining for 20 minutes and lysis for 10 minutes. Then load the sample.
Before counting, set the dilution factor to 3 as the sample is diluted by 1/3.
Reminder
Make sure to adjust the dilution factor