How do you make a 1:1000 antibody dilution?
A 1:1000 dilution is one part antibody in 1,000 parts total. Antibody volume = final volume ÷ 1,000; the rest is buffer. For 10 mL, that is 10 µL of antibody in 9.99 mL of buffer; for 5 mL, 5 µL in 4.995 mL.
How do you convert a ratio to µg/mL?
Divide the stock concentration by the dilution. A 1 mg/mL stock (1,000 µg/mL) at 1:500 is 2 µg/mL. In concentration mode, the calculator works backwards: antibody volume = working concentration x final volume ÷ stock concentration. Worked example: 2 µg/mL in 5 mL from a 1 mg/mL stock needs 10 µL of antibody, a 1:500 dilution.
Which dilution should I use?
Start from the datasheet's recommended range for your application, then titrate: run a few dilutions around it and pick the one with the best signal against background. When a volume falls under 1 µL, make an intermediate dilution first; the calculator flags it.
Frequently asked questions
What does a 1:1000 dilution mean?
One part antibody in 1,000 parts of final solution, for example 1 µL of antibody in 999 µL of buffer.
How much antibody for a 1:1000 dilution in 10 mL?
10 µL of antibody plus 9.99 mL of buffer.
How do I convert an antibody ratio to µg/mL?
Divide the stock concentration by the dilution factor. A 1 mg/mL stock at 1:1000 is 1 µg/mL.
What buffer should antibodies be diluted in?
Follow the datasheet for the application. The buffer affects background and signal, so keep it consistent between experiments.
What if the antibody volume is tiny?
Make an intermediate dilution, for example 1:10, and dilute that further, or prepare a larger final volume. Volumes under 1 µL are hard to pipette accurately.
Is anything I enter saved or sent?
No. The calculator runs in your browser. Nothing you type is saved, sent to BioBridgeX or added to the page address, and analytics never see your numbers.
Related free tools
- Dilution Calculator: C1V1 = C2V2 for any unknown, with stock and diluent volumes and the dilution factor.
- Serial Dilution Calculator: Transfer and diluent volumes and a concentration table for 2-fold, 3-fold, half-log or 10-fold series.
- Reconstitution Calculator: Solvent volume to add to a vial of lyophilized protein, antibody or compound for a target stock.
- All lab calculators and every free tool on BioBridgeX.
Sources
Check every result before use. Formulas assume ideal conditions; follow your validated lab procedures and the reagent supplier's instructions. Facts on this page last checked 27 October 2026.
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