Qproteome Bacterial Protein Prep Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Add lysozyme and Benzonase nuclease to the Qproteome Bacterial Protein Prep.	- The cell pellets are washed twice with 20 mL of washing buffer (50 m Tris‐HCI, pH 7.2), centrifuged (14 000g, 30 min at 4°C), and then disrupted by an Qproteome Bacterial Protein Prep Kit (Qiagen, Hilden, Germany) according to the Qproteome Bacterial Protein Preparation Handbook.	- The cell lysate supernatant is retained and then 100 mL of acetone are added and the samples are precipitated (12 h at −20°C). The proteins are pelleted (14 000g, 20 min) and resuspended in rehydration buffer (RB).

Upstream tips
- Add lysozyme and Benzonase nuclease to the Qproteome Bacterial Protein Prep.
Protocol tips
- The cell pellets are washed twice with 20 mL of washing buffer (50 m Tris‐HCI, pH 7.2), centrifuged (14 000g, 30 min at 4°C), and then disrupted by an Qproteome Bacterial Protein Prep Kit (Qiagen, Hilden, Germany) according to the Qproteome Bacterial Protein Preparation Handbook.
Downstream tips
- The cell lysate supernatant is retained and then 100 mL of acetone are added and the samples are precipitated (12 h at −20°C). The proteins are pelleted (14 000g, 20 min) and resuspended in rehydration buffer (RB).

Publication protocol

Proteins were extracted from three replicate bacterial cell pellets of PccS1‐V‐Ze, PccS1‐LB and PccS1‐LB‐Ze for each condition. Briefly, the cell pellets were washed twice with 20 mL of washing buffer (50 m Tris‐HCI, pH 7.2), centrifuged at 14 000 g for 30 min at 4°C, and then disrupted by an Qproteome Bacterial Protein Prep Kit (Qiagen, Hilden, Germany) according to the Qproteome Bacterial Protein Preparation Handbook. The cell pellets were thawed on ice for 15 min and resuspended in 10 mL of native lysis buffer (with lysozyme and nuclease added). The samples were incubated on ice for 30 min and then gently mixed two or three times by swirling the solution before centrifugation at 14 000 g for 30 min at 4°C to pellet the cellular debris. The cell lysate supernatant was retained and then 100 mL of acetone were added and the samples were precipitated for 12 h at −20°C. The proteins were pelleted by centrifugation at 14 000 g for 20 min and the pellet was resuspended in rehydration buffer (RB). Protein samples were stored at −20°C and protein quantification was performed according to Bradford. The concentration of the extracted protein was detected using a 2‐D Quant Kit (GE Healthcare, Chicago, IL, USA).

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 - Pectobacterium carotovorum using Qproteome Bacterial Protein Prep Kit from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for Qproteome Bacterial Protein Prep Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Protein isolation Bacteria - Pectobacterium carotovorum using Qproteome Bacterial Protein Prep Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.