Citrate Antigen Retrieval Buffer Calculator
Scale a low-pH citrate/Tween 20 antigen-retrieval buffer made with trisodium citrate dihydrate at pH 6.0.
Select a reviewed preset and enter the final volume to prepare.
| Reagent | Formula | Amount or action |
|---|---|---|
| Trisodium citrate dihydratedihydrate | 2.94 g | |
| Tween 20neat liquid | 500 µL | |
| Hydrochloric acid | Adjust experimentally to pH 6.0. | |
| Purified water | After pH adjustment, bring the total solution volume to 1 L. Do not add 1 L of water; add water only until the total solution reaches 1 L. |
How to prepare Citrate retrieval buffer pH 6.0
- Add the calculated trisodium citrate dihydrate to approximately 70–80% of the selected final volume of purified water and stir until dissolved.
- Add HCl gradually while monitoring the pH until pH 6.0 is reached.
- Add the calculated Tween 20 volume and mix gently until evenly dispersed.
- Bring to the selected final volume, mix thoroughly, and verify the final pH before use.
About This Buffer
Citrate buffer at pH 6.0 is commonly used for heat-induced antigen retrieval in immunohistochemistry. The optimal retrieval buffer and heating conditions depend on the specimen and antibody, so buffer preparation should be considered separately from the complete retrieval protocol. This calculator prepares a 10 mM trisodium citrate dihydrate formulation containing 0.05% Tween 20, with the pH adjusted experimentally to 6.0 using HCl.
Common questions
Which citrate form does this recipe use?
The preset uses trisodium citrate dihydrate. The displayed mass applies to that exact hydration state.
How much Tween 20 is in the final buffer?
The reviewed formulation contains 0.05% v/v Tween 20.
Does this page specify antigen-retrieval heating conditions?
No. It calculates the buffer recipe only. Heating time, temperature, and specimen handling should follow the validated staining protocol.
Sources and review notes
The source for the currently selected preset appears first. Recipe quantities remain tied to the cited article location.
Citrate retrieval buffer pH 6.0
Source location: Antigen Retrieval, Citrate Buffer — dissolve 2.94 g trisodium citrate dihydrate, adjust to pH 6 with HCl, add 0.5 mL Tween 20, then bring to 1 L.
Preparation method: Source-stated quantitative composition
- Immunofluorescence Labeling of Skeletal Muscle in Development, Regeneration, and Disease PMC10204082 · open article