Trizma ph 9

Manufactured by Merck Group

Sourced in United States

Trizma pH 9.0 is a buffer solution with a pH of 9.0, formulated using the Trizma buffer system. It is commonly used in various laboratory applications that require a stable alkaline pH environment.

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Lab products found in correlation

Boric acid, by Thermo Fisher Scientific (2 mentions) Amicon ultra 15 100 kda centrifugal filter devices, by Merck Group (1 mentions) Centricon plus 70 100 kda centrifugal filter devices, by Merck Group (1 mentions) Trizma, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)

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Trizma

3 protocols using trizma ph 9

Purification and Depletion of IgG from Serum

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Serum from immunized mice or rabbits was diluted 2:1 in protein G binding buffer (Pierce, UK) and applied to protein G sepharose columns. After washing twice with 3x serum volume of binding buffer, IgG was eluted in 10 mL protein G elution buffer (Pierce, UK) and restored to neutral pH by the addition of 300 μL Trizma pH 9.0 (Sigma Aldrich, USA). Where RBC pre-depletion was performed, 1 μg of packed 100% hematocrit RBC per 1 μg IgG was applied to sample, incubated at room temperature (RT) with agitation for 1 h. RBC were then pelleted by centrifugation at 1,000 xg for 5 min and the supernatant removed.

Illingworth J.J., Alanine D.G., Brown R., Marshall J.M., Bartlett H.E., Silk S.E., Labbé G.M., Quinkert D., Cho J.S., Wendler J.P., Pattinson D.J., Barfod L., Douglas A.D., Shea M.W., Wright K.E., de Cassan S.C., Higgins M.K, & Draper S.J. (2019). Functional Comparison of Blood-Stage Plasmodium falciparum Malaria Vaccine Candidate Antigens. Frontiers in Immunology, 10, 1254.

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Antigen Concentration and Inactivation

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Antigen was concentrated after inactivation, as it had been determined empirically that antigen activity was lost if it was concentrated before inactivation. Inactivated cell culture supernatants were concentrated in Amicon Ultra-15 100kDa Centrifugal Filter Devices (Millipore, Billerica, MA) or Centricon Plus-70 100-kDa Centrifugal Filter Devices (Millipore) at 3500 x g for 10-45 min at 4°C. The final volume was adjusted with 0.1M trizma/BS buffer: 1.0M trizma pH 9.0 (Sigma-Aldrich) + borate saline solution pH 9.0 [1.5M sodium chloride (Daigger, Vernon Hills, IL), 0.5M boric acid (Fisher Scientific), 1.0N sodium hydroxide (Daigger)] to the desired concentration factor.

Goodman C.H., Russell B.J., Velez J.O., Laven J.J., Bagarozzi DA J.r., Moon J.L., Bedi K, & Johnson B.W. (2015). Production of a Sindbis/Eastern Equine Encephalitis Chimeric Virus Inactivated Cell Culture Antigen. Journal of virological methods, 223, 19-24.

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Concentrating Viral Antigens for Analysis

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Inactivated cell culture supernatants were concentrated by ultracentrifugation at 54,000 x g at 4°C for ∼16 h. Supernatant was completely removed and pelleted antigen was resuspended in 0.1M trizma/BS buffer: 1.0M trizma pH 9.0 (Sigma-Aldrich) + borate saline solution pH 9.0 [1.5M sodium chloride (Daigger, Vernon Hills, IL), 0.5M boric acid (Fisher Scientific), 1.0N sodium hydroxide (Daigger)], or 1X phosphate buffered saline (PBS, Life Technologies) to achieve the desired concentration factor.

Goodman C.H., Russell B.J., Velez J.O., Laven J.J., Nicholson W.L., Bagarozzi DA J.r., Moon J.L., Bedi K, & Johnson B.W. (2014). Development of an algorithm for production of inactivated arbovirus antigens in cell culture. Journal of virological methods, 208, 66-78.

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