PBS, pH 7.4

To prepare this buffer:

(1) Make up 100 mL of a stock solution of 0.5 M EDTA by dissolving 14.6 g EDTA in approximately 80 mL of ddH2O. Adjust the pH to 7.4, then adjust the volume to 100 mL. Autoclave and filter 25 mL aliquots through a sterilized filter into 4 sterile 50 mL tubes. Store one at 4 degrees C and the rest at -20 degrees C for future use.
(2) Make up a stock solution of 20% BSA by putting 5 g of BSA in a 50 mL tube and filling it with ddH2O up to 25 mL. Leave on a roller at room temperature until dissolved. Filter through a sterile 0.2 µm filter into a sterile 50 mL tube and adjust the volume to 30 mL with sterile H2O.
(3) Make the isolation buffer by taking a 500 mL bottle of 1x PBS, pH 7.4 without Ca2+ or Mg2+ (Cat. No. 10010023). Add 2.5 mL 20% BSA and 2 mL 0.5 M EDTA. Mix by turning the bottle end over end 10 times. Store at 4 degrees C.

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This product should be stored at 2-8 degrees C and should not be frozen. The biggest issue with accidentally freezing this product is its solubility. If this product was accidentally frozen, we recommend placing it in a 2-8 degree environment and allowing it to slowly thaw overnight. If the product is fully thawed and shows no signs of precipitation, then it should still be usable, but we cannot guarantee effectiveness.

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Our PBS, pH 7.2 and PBS, pH 7.4 products are tested for endotoxin and the endotoxin levels are shown on the Certificate of Analysis (CoA) for the particular lot number of the product.

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We offer PBS - Phosphate-Buffered Saline (10X) pH 7.4, RNase-free (Cat. Nos. AM9624 and AM9625), which is molecular biology grade and is certified to be RNase-free. However, it is not sterile and hence is not cell culture-grade.

All our other PBS products (Cat. Nos. 70011xxx, 10010xxx, 70013xxx, 20012xxx) are filter-sterilized using a 0.1 µm filter and are cell culture grade (ready to use directly on cells). However, they are not tested for nuclease activity.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

We offer PBS - Phosphate-Buffered Saline (10X) pH 7.4, RNase-free (Cat. Nos. AM9624 and AM9625) that is molecular biology grade and is certified to be RNase-free. Our other PBS products are made with distilled water and are filter-sterilized, but are not tested for RNase and DNase activity.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.