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| Product Name | Fixation Buffer for Flow Cytometry |
| Catalog No. | RESK-0327 |
| Description | This Fixation Buffer is designed for treating cell membranes prior to intracellular cytokine staining, together with a permeabilization buffer. The fixation step stabilizes the cell membrane and preserves the binding of surface antibodies to their antigens, while the subsequent permeabilization step creates small pores in the intact cell membrane to allow antibodies to enter the cell. After fixation and permeabilization of the test cells, the corresponding antibodies or detection reagents are added, and following incubation the samples can be analyzed directly by flow cytometry for intracellular antigens and DNA. |
| Intended Use | Fixation of cells prior to intracellular antigen, cytokine, or DNA detection by flow cytometry. |
| Principle / Technology | The fixative stabilizes the cell membrane and cross-links cellular proteins, preserving cell morphology and surface antibody-antigen binding; combined with a permeabilization reagent, it enables antibodies to access intracellular targets for flow cytometric analysis. |
| Detection Method | Flow cytometry |
| Sample Type | Single-cell suspensions (e.g., cultured cells) for intracellular staining |
| Reaction Conditions / Protocol | 1. Harvest the test cells and centrifuge at 1000 rpm for 10 min; discard the supernatant. 2. Add 500 uL of pre-cooled (4 °C) Fixation Buffer. 3. Incubate at room temperature for 10 min. 4. Centrifuge at 1000 rpm for 10 min. 5. Discard the supernatant, wash once with PBS, and centrifuge at 1000 rpm for 10 min. 6. Discard the supernatant and add 1.5 mL of Permeabilization Buffer. 7. Incubate at room temperature for 10-15 min. 8. Centrifuge at 1000 rpm for 5 min. 9. Discard the supernatant and resuspend the cells in an appropriate volume of Permeabilization Buffer. 10. Add the detection antibody for staining, then analyze by flow cytometry. |
| Components / Formulation | Ready-to-use fixation solution; a 4% paraformaldehyde formulation is referenced for this fixation application. |
| Storage Conditions | Store at 2-8 °C protected from light. Do not freeze. |
| Shelf Life | 1 year |
| Package Specifications | 50 mL |
| Product Form | Liquid |
| Regulatory / Compliance | For research use only. Not for use in diagnostic or therapeutic procedures. |
| Compatibility | Used in combination with a permeabilization wash buffer and downstream antibody staining for flow cytometric analysis of intracellular antigens and DNA. |
| Recommended Buffer System | PBS for washing; Permeabilization Buffer for the permeabilization and antibody-staining steps. |
| Application Notes / Precautions | Use pre-cooled (4 °C) Fixation Buffer for cell fixation. Protect from light and do not freeze. Fixation should be followed by permeabilization before adding antibodies against intracellular targets. |
For research use only, not for clinical use.
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