Gyros IncSupplier Diversity Partner REXXIP HN BUFFER 25 ML PER VI DFS Item

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- There is a previous cell lysate step.	- Total cell lysate is subjected to ultracentrifugation (435,700 × g, 1 h, 4°C) to pellet membrane vesicles. - Membrane pellets are rinsed in 1.0 ml of HN buffer with protease inhibitors, reisolated by ultracentrifugation, and suspended in 200 to 400 μl of HN buffer plus complete protease inhibitor cocktail with the aid of a coated homogenizer.	- Protein precipitation is performed following Trichloroacetic Acid Solution, MP Biomedicals™ protocol (Fisher Scientific).

Upstream tips
- There is a previous cell lysate step.
Protocol tips
- Total cell lysate is subjected to ultracentrifugation (435,700 × g, 1 h, 4°C) to pellet membrane vesicles. - Membrane pellets are rinsed in 1.0 ml of HN buffer with protease inhibitors, reisolated by ultracentrifugation, and suspended in 200 to 400 μl of HN buffer plus complete protease inhibitor cocktail with the aid of a coated homogenizer.
Downstream tips
- Protein precipitation is performed following Trichloroacetic Acid Solution, MP Biomedicals™ protocol (Fisher Scientific).

Publication protocol

Total cell lysate was subjected to ultracentrifugation (435,700 × g, 1 h, 4°C) in a Sorvall Discovery M150 SE Ultracentrifuge to pellet membrane vesicles. Membrane pellets were rinsed in 1.0 ml of HN buffer with the addition of protease inhibitors, reisolated by ultracentrifugation, and suspended in 200 to 400 μl of HN buffer plus complete protease inhibitor cocktail with the aid of a small Teflon coated homogenizer (Kontes, Vineland, NJ). Suspended membrane-associated proteins were stored at −80°C until use. Membrane protein quantities were determined by using a modified Lowry protein assay with bovine serum albumin as a standard.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein isolation Bacteria - Borrelia burgdorferi using Gyros IncSupplier Diversity Partner REXXIP HN BUFFER 25 ML PER VI DFS Item from Fisher Scientific.

Manufacturer protocol

Download the product protocol from Fisher Scientific for Gyros IncSupplier Diversity Partner REXXIP HN BUFFER 25 ML PER VI DFS Item below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Protein isolation Bacteria - Borrelia burgdorferi using Gyros IncSupplier Diversity Partner REXXIP HN BUFFER 25 ML PER VI DFS Item from Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.