Carbonate/Bicarbonate Buffer(CBB) Preparation and Protocol (PH: 9.6)


Carbonate/Bicarbonate Buffer(CBB) Preparation and Protocol is useful for Immunoassay coating, Bio-conjugation and biochemistry methods. Sodium Carbonate Bicarbonate Buffer used in a range of protein conjugation procedures, protein and antibody conjugation, immunoassay applications, biology, and biochemistry methods. It is also a proper buffered solution for alkaline phosphatase Immunoassay systems. In combination of Borate, Glycine or Ammonia, CBB buffer is perfect for Biochemical applications. CBB has good buffering capacity and is easy to prepare; it is proper buffer just for Basic PH in a range from 8.2-10.0, because it is PH sensitive and hydrolyses to CO2 and H2O. Even for mentioned Basic PH, CBB needs some stabilizers to extend its shelf life. Normally, CBB should be prepared and be used freshly.

The most common use of Carbonate Bicarbonate Buffer (CBB) is ELISA Coating Solution of Abs in Immunoassy methods in PH=9.6. Carbonate Bicarbonate Buffer is not suitable for Ag coating in Direct and Indirect ELISA, coating of recombinant proteins, and non-Glycolyzed proteins such as Streptavidin. In these cases, CBB needs additional stabilizers to keep proteins stable during the coating time. However, it can be used for many non-commercial ELISA coating in research labs!

Sodium Carbonate bicarbonate buffer is an affordable buffer solution and nontoxic component that make it a green Buffer for the environment. It is sensitive to direct sunlight, temperature and microbial contamination and needs to be kept in closed containers.

Carbonate Bicarbonate buffer preparation (PH: 9.6)

Recommended Concentration for CBB is 100 mM solution and ready to use form.

1. For a 0.1 M solution Sodium Carbonate Bicarbonate Buffer, dissolve 1.59 g of Na2CO3 and 2.93gr NaHCO3 in 950 mL of distilled water.

                               2.  Adjust the pH to 9.6 value by slowly adding approximately HCl.

                     3. Adjust the volume of the solution to 1000 mL with distilled water.

4. If necessary, Filter Tris buffer with 0.22 μm sterile filter.

Note1: Add slowly with shaking the buffer. CBB is sensitive to rapid PH change. Be cautious of making any bubble upon adding HCl.

Note2: For Immunoassay tests, you do not need to sterilize CBB.

Note3: CBB buffer is stable at 4C° at least for 4 days.

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