CulGenex™

Phosphate Buffered Saline Powders

Cat. no. C1608 CulGenex™ Phosphate Buffered Saline (PBS), 1X, pH 7.3, 1L Pouch 6 pouches/pack
Cat. no. C1618 CulGenex™ Phosphate Buffered Saline, Dulbecco's (DPBS), 1X, pH 7.2, 1L Pouch 6 pouches/pack
Cat. no. C1638 CulGenex™ Phosphate Buffered Saline (PBS), 1X, pH 7.6, 1L Pouch 6 pouches/pack
Cat. no. C1648 CulGenex™ Phosphate Buffered Saline, Dulbecco's (DPBS), with Ca++ and Mg++, 1X, pH 7.4, 1L Pouch 6 pouches/pack

INTENDED USE

Hardy Diagnostics CulGenex™ Phosphate Buffered Saline Powders are recommended for use in various procedures used in biological and molecular research. (1-5)

This dehydrated culture medium is a raw material intended to be used in the making of prepared media products, which will require further processing, additional ingredients, or supplements.

SUMMARY

Commonly used in biological and biomedical research, Phosphate Buffered Saline (PBS) has many applications due to its isotonic nature. PBS can be used as a diluting agent in preparing decimal dilutions and as a rinsing agent for rinsing labware containing cells. Phosphate salts are nontoxic to living cells and have a high buffering capacity; pH maintenance is important in retaining cell viability and in the recovery and revitalization of injured or damaged cells during bacterial culture and analysis. Phosphate Buffered Saline contains sodium chloride, sodium phosphate, potassium phosphate and, in some formulations, potassium chloride, magnesium and calcium.

Dulbecco's Phosphate Buffered Saline (DPBS) is commonly used to provide a buffer system to maintain cell culture media in the physiological pH range of 7.0 to 7.6, where a chemically defined balanced salt solution is needed to maintain cell tonicity and viability in vitro . DPBS is also commonly used in cell enumerations as a diluting, irrigating or transporting fluid, and as a buffer in many chromotographic procedures. Hardy Diagnostics CulGenex™ DPBS buffer is available with or without the addition of calcium and magnesium ions.

Hardy Diagnostics CulGenex™ Phosphate Buffered Saline Powders are packaged in a pouch that will make one liter upon the addition of water. The product is ready to mix at the 1X concentration for convenient use. They are available in packages of six.

FORMULA*

Phosphate Buffered Saline (PBS), 1X (Cat. no. C1608):
Gram weight per liter: 9.9gm/L
Sodium Chloride 9.0gm
Disodium Phosphate 0.795gm
Monopotassium Phosphate 0.144gm

Final pH 7.3 +/- 0.1 at 25ºC.


Phosphate Buffered Saline, Dulbecco's (DPBS), 1X (Cat. no. C1618):
Gram weight per liter: 10.6gm/L
Sodium Chloride 8.0gm
Disodium Phosphate 2.16gm
Potassium Chloride 0.2gm
Monopotassium Phosphate 0.2gm

Final pH 7.2 +/- 0.1 at 25ºC.


Phosphate Buffered Saline (PBS), 1X (Cat. no. C1638):
Gram weight per liter: 10.0gm/L
Sodium Chloride 9.0gm
Disodium Phosphate 0.795gm
Monopotassium Phosphate 0.144gm

Final pH 7.6 +/- 0.1 at 25ºC.


Phosphate Buffered Saline, Dulbecco's (DPBS), with Ca++ and Mg++, 1X (Cat. no. C1648):
Gram weight per liter: 9.8gm/L
Sodium Chloride 8.0gm
Disodium Phosphate 1.15gm
Potassium Chloride 0.2gm
Monopotassium Phosphate 0.2gm
Calcium Chloride 0.1gm
Magnesium Chloride 0.1gm

Final pH 7.4 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Store the sealed container(s) holding dehydrated powder at 2-30ºC. Dehydrated powder is very hygroscopic. Keep lid tightly sealed. Protect dehydrated powder from moisture and light. The dehydrated powder should be discarded if it is not free-flowing or if the color has changed from its original white.

Store the prepared saline at 2-30ºC.


PRECAUTIONS


METHOD OF PREPARATION FOR DEHYDRATED BUFFERS

1. Empty the contents of one packet into one liter of deionized water, DEPC treated water (Cat. no. CG482, CG482BX), or ultra pure molecular grade water (Cat. no. CG492, CG492BX). Stir to mix thoroughly.

2. Filter sterilize using a 0.1um filter.

PROCEDURE

General Dilution Guidelines:

1:10 Serial Dilutions
1. Using a sterile pipet, aliquot 10ml of test suspension to 90ml of PBS diluent. Mix thoroughly. This yields a 1:10 dilution.

2. Use a second sterile pipet to aliquot 10ml of the 1:10 dilution prepared in step 1 into a second 90ml filled vessel of PBS diluent. Mix thoroughly. This yields a 1:100 dilution.

3. Continue aliquoting 10ml of subsequent dilutions into 90ml filled PBS diluent vessels until the desired concentration of test sample is obtained. Each succeeding dilution increases by a factor of 10. A separate sterile pipet should be used with each dilution.

1:100 Serial Dilutions
1. Using a sterile pipet, aliquot 1ml of test suspension to 99ml of PBS diluent. Mix thoroughly. This yields a 1:100 dilution.

2. Use a second sterile pipet to aliquot 1ml of the 1:100 dilution prepared in step 1a into a second 99ml filled vessel of PBS diluent. Mix thoroughly. This yields a 1:10,000 dilution.

3. Continue aliquoting 1ml of subsequent dilutions into 99ml filled PBS diluent vessels until the desired concentration of test sample is obtained. Each succeeding dilution increases by a factor of 100. A separate sterile pipet should be used with each dilution.

Consult additional references for information on specific standard protocols. (1-5)

For information on procedures and interpretation of results, consult listed references.

LIMITATIONS

Note: PBS with calcium and magnesium is not recommended for enzymatic reactions due to its potential interaction with divalent cations and resulting precipitation.


MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological and molecular biology supplies and equipment such as autoclaves, incinerators, and incubators, deionized water, DEPC treated water (Cat. no. CG482, CG482BX), or ultra pure molecular grade water (Cat. no. CG492, CG492BX), etc., are not provided.

QUALITY CONTROL

Hardy Diagnostics tests CulGenex™ Phosphate Buffered Salines (PBS), 1X; Phosphate Buffered Saline, Dulbecco's (DPBS), 1X; and Phosphate Buffered Saline, Dulbecco's (DPBS), with Ca++ and Mg++, 1X for osmolality and pH.

User Quality Control


Physical Appearance

CulGenex™ Phosphate Buffered Saline (PBS), 1X, pH 7.3; Phosphate Buffered Saline (PBS), 1X, pH 7.6; Phosphate Buffered Saline, Dulbecco's (DPBS), 1X, pH 7.2; and Phosphate Buffered Saline, Dulbecco's (DPBS), with Ca++ and Mg++, 1X, pH 7.4 powder should appear homogeneous, free-flowing, and white in color. The prepared media should appear clear, and colorless.

REFERENCES

1. Ausubel, F.M., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, K. Struhl, Editors. 2010. Current Protocols in Molecular Biology . John Wiley and Sons, Inc. Malden, MA.

2. Bullock, G.R. and Petrusz, P. 1989. Techniques in Immunocytochemistry , Volumes 1, 2, 3 and 4, Academic Press, London.

3. Cseke, L.J., P.B. Kaufman, G.K. Podila, and C.J. Tsai. 2004. Handbook of Molecular and Cellular Methods in Biology and Medicine . CRC Press.Taylor & Francis LLC. Boca Raton, FL.

4. Sambrook and Russell. 2001. Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press. Woodbury, New York.

5. Walker, J.M. 1984. Methods in Molecular Biology . The Humana Press Inc. Clifton, NJ.


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