Production of His-tagged or Biotinylated Salipro-PANX1

To generate His-tagged or biotinylated Salipro-PANX1 particles, the same method as described above was used to extract and purify Salipro-PANX1 using His-tagged or biotinylated saposin A. His-tagged saposin A was produced and purified as previously described^{3 (link),6}, albeit without the TEV protease cleavage step for His-tag removal. Saposin A was biotinylated in a one-hour incubation step at RT using NHS-PEG12-Biotin (Thermo Fisher Scientific), following the manufacturer’s recommendations. The Quant*Tag™ Biotin Kit (Vector Laboratories) was used to confirm the amount of saposin A biotinylation.

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Drulyte I., Gutgsell A.R., Lloris-Garcerá P., Liss M., Geschwindner S., Radjainia M., Frauenfeld J, & Löving R. (2023). Direct cell extraction of membrane proteins for structure–function analysis. Scientific Reports, 13, 1420.

Publication 2023

NHS-PEG12-Biotin Quant*Tag™ Biotin Kit

Biotin Biotinylation Cleavage Nhs biotin Saposin a Step one Tev protease Vector

Corresponding Organization :

Other organizations : Thermo Fisher Scientific (Netherlands), AstraZeneca (Sweden), Active Biotech (Sweden), Thermo Fisher Scientific (Germany)

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Variable analysis

independent variables

Use of His-tagged saposin A
Use of biotinylated saposin A

dependent variables

Extraction and purification of Salipro-PANX1 particles

control variables

The same method as described above was used to extract and purify Salipro-PANX1

positive controls

His-tagged saposin A production and purification as previously described

negative controls

No negative controls were explicitly mentioned

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