How It Works
This calculator works in 4 steps.
Enter the primary dilution
Type the protocol ratio, such as 1:1,000.
Enter the volume per unit
Type the incubation volume per blot, slide, or well.
Enter the count and overage
Type the number of units and the overage percent.
Read the batch volumes
You get the total antibody and buffer volumes.
The Formula
The formula multiplies the volume per unit by the count and the extra. Then it divides by the ratio.
V_total = V_unit × N × (1 + o)V_primary = V_total / X- V_unit:
- Incubation volume per unit
- N:
- Number of blots, slides, or wells
- o:
- Overage fraction
- X:
- Ratio term in 1:X
Three mini-blots at 5 mL each with 1:1,000 primary need 15 mL working solution: 15 µL antibody plus 14,985 µL blocking buffer.
Conversion Table
The conversion table gives antibody and diluent volumes for 6 primary dilutions and working volumes.
| Primary dilution | Working volume | Primary antibody | Diluent |
|---|---|---|---|
| 1:500 | 5 mL | 10 µL | 4,990 µL |
| 1:1,000 | 10 mL | 10 µL | 9,990 µL |
| 1:1,000 | 15 mL | 15 µL | 14,985 µL |
| 1:2,000 | 10 mL | 5 µL | 9,995 µL |
| 1:100 | 0.5 mL | 5 µL | 495 µL |
| 1:200 | 1 mL | 5 µL | 995 µL |
What Is the Primary Antibody Dilution Calculator?
The Primary Antibody Dilution Calculator is a tool that turns a primary antibody ratio into volumes for a blot, a set of slides or a plate. The primary binds the target protein. So its dilution sets how clean the signal is.
The volume you need depends on the format. A mini-blot in a box uses 5 to 10 mL. A sealed bag uses 2 to 3 mL. A slide uses 100 to 200 µL per section. The calculator multiplies the volume per blot or slide by the count. It adds a little extra, then gives the antibody volume for the batch.
Common Uses
There are 5 main uses of this calculator.
Multi-blot batches
Users prepare 1 working solution for several blots.
Slide staining runs
IHC labs scale primary volumes to the slide count.
Titration grids
Researchers test 1:500, 1:1,000, and 1:2,000 in parallel.
Antibody budgeting
Labs estimate how many experiments a 100 µL vial supports.
Protocol records
Users log exact primary volumes for repeat runs.