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| Product Name | Phosphate-Buffered Saline (PBS), 10x Concentrate, pH 7.4, Sterile, Cell Culture Grade |
| Catalog No. | ATR-B-YJL-0200 |
| Description | 10x concentrated phosphate-buffered saline (PBS) designed for dilution to 1x isotonic working solution for cell culture, immunoassays, and general biochemical applications. The 1x solution (after 1:10 dilution with ultrapure water) yields 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 1.8 mM KH2PO4 at pH 7.4, providing an osmotically balanced, non-toxic isotonic buffer (approximately 300 mOsm/L) compatible with mammalian cells. This formulation contains no calcium or magnesium (DPBS without Ca2+/Mg2+), making it ideal for cell washing prior to trypsinization and for applications where divalent cations may interfere, such as immunoprecipitation and some enzyme assays. The buffer is 0.1 um sterile-filtered and tested for the absence of endotoxin, mycoplasma, DNase, RNase, and protease. |
| Intended Use | Isotonic buffer for: washing mammalian cells before trypsinization or harvesting; diluting cells for counting and plating; rinsing cell monolayers; diluting antibodies and proteins for immunoassays (ELISA, western blot, immunohistochemistry); protein and antibody conjugation reactions; tissue rinsing in histology; preparation of working solutions for biochemical assays; and general laboratory use as a physiological saline buffer. |
| Principle / Technology | PBS maintains physiological pH (7.4) and osmolarity (approximately 300 mOsm/L) through the phosphate buffer system (H2PO4-/HPO4^2-, pKa 7.2 at 25 C). NaCl provides the bulk osmolarity and ionic strength mimicking extracellular fluid. KCl provides K+ ions at physiological intracellular concentrations. The absence of Ca2+ and Mg2+ prevents: cell clumping during monolayer washing; activation of integrins and cadherins; precipitation of phosphate as insoluble calcium/magnesium phosphate; and interference with EDTA-based cell detachment. |
| Detection Method | Dilute 10x PBS 1:10 with ultrapure (cell culture grade) water to prepare 1x PBS (137 mM NaCl, 2.7 mM KCl, 10 mM phosphate, pH 7.4). Mix thoroughly. If absolute sterility is required for cell culture, use in a biosafety cabinet with aseptic technique after opening. Warm 1x PBS to 37 C before use with live cells to prevent thermal shock. 1x PBS stored at RT or 2-8 C should be monitored for microbial contamination. |
| Sample Type | 1x working solution (1:10 dilution of 10x concentrate) used for cell washing, sample dilution, and assay buffer preparation. |
| Performance Range / Specifications | 1x working concentration: 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4 +/-0.1; osmolarity (1x): 280-320 mOsm/L; conductivity (1x): 14-16 mS/cm at 25 C; endotoxin: <0.05 EU/mL; sterility: no growth after 14-day incubation (bacterial, fungal). |
| Sensitivity / LOD | Not applicable — buffer solution. |
| Specificity | PBS is a general-purpose physiological buffer; phosphate buffer system has a pKa of 7.2, providing optimal buffering at pH 7.0-7.6. Not to be confused with DPBS containing Ca2+/Mg2+ (used for adherent cell maintenance) or Tris-buffered saline (TBS, used for immunoassays where phosphate may interfere with alkaline phosphatase detection). |
| Reaction Conditions / Protocol | Buffer is physiologically compatible with mammalian cells at 37 C; 1x PBS used as washing and dilution buffer in cell culture, immunoassays, and biochemical reactions. |
| Components / Formulation | 10x PBS Concentrate (1 L): 1.37 M NaCl (80.1 g/L), 27 mM KCl (2.01 g/L), 100 mM Na2HPO4 (anhydrous, 14.2 g/L or dodecahydrate, 35.8 g/L), 18 mM KH2PO4 (2.45 g/L), pH 7.4 +/-0.1 (adjusted with HCl or NaOH), 0.1 um sterile-filtered, no calcium, no magnesium, no preservatives. |
| Storage Conditions | Store at RT (15-25 C); protect from contamination — use aseptic technique when pouring or pipetting; 10x may precipitate at temperatures below 10 C; if precipitation occurs, warm to 37 C and mix until dissolved. |
| Shelf Life | 24 months from date of manufacture at RT; after dilution to 1x: 1 month at RT aseptic, 3 months at 2-8 C. |
| Package Specifications | 500 mL (makes 5 L 1x), 1 L (makes 10 L 1x), 4 L (makes 40 L 1x). |
| Product Form | Clear, colorless liquid; 10x concentrate; may form slight precipitate at cold temperatures (reversible). |
| Quality Control | Each lot: pH 7.3-7.5 (1x, 25 C); osmolarity 280-320 mOsm/L (1x); conductivity 14-16 mS/cm (1x); sterility: bacterial and fungal count <0.1 CFU/mL, no growth 14-day test; endotoxin: <0.05 EU/mL; mycoplasma: negative (PCR); DNase/RNase: not detected; cell culture test: >=95% viability of HEK293T and HeLa cells after 2-hour incubation in 1x PBS at 37 C. |
| Key Features | 10x concentrate for convenience and storage; 0.1 um sterile-filtered; cell culture grade; calcium-free and magnesium-free; low endotoxin (<0.05 EU/mL); mycoplasma tested negative; 24-month shelf life; no preservatives. |
| Purity | All components: cell culture grade or higher; NaCl, KCl, Na2HPO4, KH2PO4: ACS grade, >=99% purity; water: cell culture grade (WFI quality, 18.2 MOhm-cm); no detectable heavy metals. |
| Concentration | 10x concentrate: 1.37 M NaCl, 27 mM KCl, 100 mM Na2HPO4, 18 mM KH2PO4; 1x working: 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4. |
| Activity / Unit Definition | Not applicable — buffer solution. |
| Molecular Weight | NaCl: 58.44 g/mol; KCl: 74.55 g/mol; Na2HPO4: 141.96 g/mol (anhydrous); KH2PO4: 136.09 g/mol. |
| Source / Origin | All chemicals: synthetic (inorganic salt synthesis); water: ultrapure deionized WFI quality; no animal-derived components; no human-derived components. |
| pH Range / Optimal pH | 1x pH 7.4 +/-0.1; phosphate pKa2 = 7.2 at 25 C; effective buffering range pH 6.5-7.9; pH decreases approximately 0.03 per degree Celsius temperature increase. |
| Shipping Conditions | Ambient temperature. |
| Expiration Date / Stability | 24 months at RT unopened; after dilution to 1x: 1 month at RT (aseptic handling), 3 months at 2-8 C; do not freeze 10x PBS (salt precipitation may be irreversible). |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic use. Not for human or animal injection without further processing. Cell culture grade but not GMP-certified for clinical manufacturing. |
| Compatibility | Compatible with all mammalian cell lines (adherent and suspension). Ca2+/Mg2+-free formulation is compatible with trypsin-EDTA-based cell detachment (EDTA chelates Ca2+/Mg2+ required for cadherin-mediated cell adhesion). Compatible with most antibodies, enzymes, and proteins in short-term incubations. Compatible with ELISA, western blot, and immunohistochemistry blocking and washing steps. Not compatible with: assays requiring Ca2+ or Mg2+ (add these separately if needed); alkaline phosphatase detection (phosphate in PBS competes with ALP substrate, use TBS instead); long-term protein storage (PBS lacks cryoprotectants and antimicrobial agents). |
| Recommended Buffer System | 10x PBS: 1.37 M NaCl, 27 mM KCl, 100 mM Na2HPO4, 18 mM KH2PO4, pH 7.4; 1x PBS: 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4. |
| Application Notes / Precautions | If 10x PBS forms a precipitate, warm to 37 C in a water bath and agitate until fully dissolved before diluting. Always dilute 10x PBS with cell culture grade water to avoid endotoxin and nuclease contamination of the 1x solution. When adding CaCl2 and/or MgCl2 to 1x PBS for specific applications (e.g., DPBS with Ca2+/Mg2+), add from sterile concentrated stocks (1 M) after dilution to 1x — adding Ca2+/Mg2+ to 10x PBS will cause precipitation of calcium/magnesium phosphate. For cell washing: use 5-10 mL 1x PBS per T75 flask, gently rock to rinse monolayer, aspirate, and repeat if needed. For immunoassays: 1x PBS with 0.05% Tween-20 (PBST) is a standard wash buffer. |
| Batch-to-Batch Consistency | pH 7.3-7.5 for every lot; osmolarity 280-320 mOsm/L; sterility and endotoxin <0.05 EU/mL confirmed for each batch. |
For research use only, not for clinical use.
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