Tris Buffered Saline (TBS), pH 8.0±0.2, is a 10X Tris-NaCl buffer concentrate containing 0.5M Tris-HCl and 1.5M sodium chloride. Dilute one volume of concentrate to a final volume of ten volumes with deionized water to prepare 1X TBS containing 50mM Tris-HCl and 150mM sodium chloride.
TBS is commonly used as a wash buffer and antibody diluent in immunochemical methods, including western blotting and ELISA. It is also used with antigen retrieval solutions in histology workflows. Because the pH of Tris buffers varies with temperature and concentration, verify the pH of the diluted buffer at the intended working temperature when tight pH control is required.
Key Properties
- Product number: 24082
- Format: 10X liquid concentrate
- pH: 8.0±0.2, approximate at 25°C
- 10X Tris-HCl concentration: 0.5M
- 10X sodium chloride concentration: 1.5M
- 1X Tris-HCl concentration after dilution: 50mM
- 1X sodium chloride concentration after dilution: 150mM
- Dilution: 1 volume concentrate to 10 final volumes
- Available sizes: 500mL and 1L
- Storage: 4°C
Applications
Western Blotting
TBS is commonly used for membrane washing and antibody dilution in western blotting. When a method specifies TBS with Tween-20 (TBST), add detergent to the diluted buffer at the concentration specified by the protocol.
ELISA & Immunoassays
Tris-NaCl buffered saline is used as a wash buffer and antibody diluent in ELISA and related immunoassays. Detergent concentration, blocking reagent, antibody concentration, and wash conditions should be selected for the specific assay.
Antigen Retrieval Workflows
TBS is used as a buffered saline with antigen retrieval solutions in histology and immunohistochemistry workflows. For a related reagent, see the Citrate Buffer Kit, pH 6.0, for antigen retrieval.
Buffer Composition & Method Considerations
- 10X composition: 0.5M Tris-HCl and 1.5M sodium chloride
- 1X composition after ten-fold dilution: 50mM Tris-HCl and 150mM sodium chloride
- The stated pH is approximate at 25°C
- Tris-buffer pH varies with both temperature and concentration
- TBS and TBST are distinct buffer systems; detergent is added when required by the experimental protocol
- TBS is phosphate-free and is commonly selected for alkaline phosphatase detection systems because phosphate can inhibit alkaline phosphatase activity
For pH-sensitive procedures, prepare the 1X working buffer, equilibrate it to the intended operating temperature, and verify the pH before use. If adjustment is required, HCl or NaOH may be used as appropriate for the method.
Application Tips
- Dilute one volume of 10X concentrate to a final volume of ten volumes with deionized water
- To prepare 1L of 1X TBS, add 100mL of 10X concentrate and bring to a final volume of 1L with deionized water
- Mix thoroughly after dilution
- For pH-sensitive assays, verify the pH after dilution at the intended working temperature
- When preparing TBST, add detergent only at the concentration specified by the experimental protocol
- Keep buffer dilution, pH, temperature, detergent concentration, and wash conditions consistent when comparing experimental samples
Handling & Storage
- Store the 10X concentrate at 4°C
- Keep the container closed when not in use
- Use deionized water for preparation of the working dilution
- Use clean laboratory vessels and measuring equipment to minimize contamination of the working buffer
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FAQ
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1. What is the composition of 10X TBS?
The 10X concentrate contains 0.5M Tris-HCl and 1.5M sodium chloride.
2. What is the composition of 1X TBS after dilution?
A ten-fold dilution gives 1X TBS containing 50mM Tris-HCl and 150mM sodium chloride.
3. How do I dilute 10X TBS to 1X?
Dilute one volume of concentrate to a final volume of ten volumes with deionized water. To prepare 1L of 1X TBS, use 100mL of 10X concentrate and bring to a final volume of 1L with deionized water.
4. What is the pH of this TBS buffer?
The stated pH is 8.0±0.2 and is approximate at 25°C. Because Tris pH depends on temperature and concentration, verify the diluted buffer at the intended working temperature when exact pH control is required.
5. Is TBS the same as TBST?
No. TBS is a Tris-NaCl buffered saline. TBST is TBS containing a nonionic detergent, commonly Tween-20. Add detergent according to the experimental protocol when TBST is required.
6. Can TBS be used for western blotting?
Yes. TBS is commonly used for membrane washing and antibody dilution in western blotting. Some protocols specify TBST rather than TBS alone.
7. Can TBS be used for ELISA?
Yes. TBS is used as a wash buffer and antibody diluent in ELISA and other immunoassays. The appropriate detergent, blocking reagent, and wash conditions depend on the assay.
8. Why is TBS used with alkaline phosphatase detection systems?
TBS is phosphate-free. Phosphate can inhibit alkaline phosphatase activity, so Tris-based saline buffers are commonly selected for alkaline phosphatase immunodetection methods.
9. Can this TBS be used with antigen retrieval solutions?
Yes. TBS is used as a buffered saline with antigen retrieval solutions in histology workflows.
10. How should the 10X concentrate be stored?
Store at 4°C.
11. What sizes are available?
Tris Buffered Saline Cat. #24082 is available in 500mL and 1L sizes. Catalog number 24082-1 is the 1L format.
12. Can larger quantities be requested?
For quantities beyond the standard offering, use the bulk-quote option.
Safety & Documentation
Refer to Technical Data Sheet #526 for buffer composition, dilution, and pH information and the Safety Data Sheet for handling and storage information. Additional formulations are available in the Life Sciences Buffers collection.
