BIS-TRIS

Basic information:

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• CAS No.: 6976-37-0


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• Useful pH range: 5.8 – 7.2








 
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Application of BIS-TRIS:

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• In the analysis of purity and genome integrity of hemophilia B gene therapy with AAV8 vector and the determination of biological efficacy, It is used as a separation colloid in the analysis step of gel electrophoresis. ^[2]








 
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HEPES

Basic information:

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• CAS No.: 15471-17-7


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• Useful pH range: 6.8 – 8.2








 
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Application of HEPES:

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• Used in the production and purification of AAV vectors as the buffer of the acidic lysis buffer for HEK293T cells to neutralizing pH value.^[1]


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• For AAV production, It was used as a buffer for the cell culture of HEK293T to maintain the pH value of the cell culture media. According to the experimental results, it significantly increases the viral plasmids release and virus production. ^[1]








 
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If you need to prepare the cell culture media of the special cell line,

we also provide HEPES (cell culture grade) that has passed the cytotoxicity test.

Further reading:Why use HEPES?

TRIS

Basic information:

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• CAS No.: 77-86-1


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• Useful pH range: 7.2 – 9.0








 
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Application of TRIS:

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• In the process of purification of lentiviral vectors, because the PBS buffer lacks buffering capacity at lower temperatures and is incompatible with the Benzonase step, TRIS is the best choice as a detergent here. In addition, TRIS can also avoid any pH restriction in the virus binding to the AEXc matrix.^[3]


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• In process of purification of Viral Particles, Use the Cells harvested by the process of Viral Particle Production were resuspended in lysis buffer (50 mM Tris, 150 mM NaCl, 2 mM MgCl2, pH 7.5). Then viral particles were released from cells with three freeze-thaw cycles.^[5]








 
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TRIS-HCL

Basic information:

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• CAS No.: 1185-53-1


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• Useful pH range: 7.2 – 9.0








 
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Application of TRIS-HCL:

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• In the analysis of purity and genome integrity of hemophilia B gene therapy with AAV8 vector and the determination of biological efficacy, It is used as the incubation solution after the gel electrophoresis analysis step. ^[2]








 
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Further reading:How to prepare adeno-associated viruses (AAV) vectors?

CHAPS

Basic Information:

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• CAS No.: 75621-03-3








 
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Application of CHAPS:

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• For proteomics: In TMT sample preparation for mammalian cell and tissue analysis, using CHAPS lysis buffer can avoid accidental protein degradation during protein digestion and improve protein extraction efficiency.^[4]


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• The remaining DNA contamination was degraded by incubation with benzonase nuclease at 37C prior to the addition of CHAPS (0.5% w/v). The crude lysate was cleared from cell debris by centrifugation (3000 g, 10 min). ^[5]


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• CHAPS has been successfully used to dissolve internal membrane proteins and receptors and has the ability to maintain protein activity.^[6]


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• Proteins can be iodinated in the presence of CHAPS. ^[6]








 
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Further reading: What is gene therapy?

Further reading: The four kinds of viral vectors which most use in gene therapy

Further reading:Why use CHAPS?

Reference:

^[1] Production of adeno-associated virus vectors for in vitro and in vivo applications: https://www.nature.com/articles/s41598-019-49624-w

^[2] Development of an In Vitro Biopotency Assay for an AAV8 Hemophilia B Gene Therapy Vector Suitable for Clinical Product Release: https://www.sciencedirect.com/science/article/pii/S2329050120300437

^[3] Downstream Processing of Lentiviral Vectors: Releasing Bottlenecks https://www.liebertpub.com/doi/pdf/10.1089/hgtb.2012.059

^[4] TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7442215/

^[5] ijms-20-05702.pdf

^[6] Method for the production and purification of adenoviral vectors: https://patents.google.com/patent/US7732129B1/en