B-PER™ Bacterial Protein Extraction Reagent

Protein isolation Bacteria - Synechocystis sp (6803)_Cyanobacteria

Experiment
Protein isolation Bacteria - Synechocystis sp (6803)_Cyanobacteria
Product
B-PER™ Bacterial Protein Extraction Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
- Proteins are collected by following the B-PER Bacterial Protein Extraction Reagent manufacturer protocol.

Publication protocol

Fifty milliliters of an exponential-phase Synechocystis PCC 6803 culture was collected. Following the B-PER and control treatments, cell pellets and medium supernatants were diluted in the ratio of 3:1 (v/v) in 4× Laemmli sample buffer and loaded in equal proportion on a 12% (v/v) Criterion™ TGX Stain-Free™ Protein Gel, which contains trihalo compounds that can react with tryptophan residues under the UV irradiation and gives rise to fluorescence (BioRad, US). Proteins were subsequently transferred from the gel to a polyvinylidene difluoride membrane. Immunodetection of YFP was carried out using anti-GFP mouse IgG (Roche Applied Science, Germany) according to the manufacturer instructions. Relative signal was quantified using Image Lab™ Software (BioRad, US). The total proteins in the gels were visualized under the UV irradiation according to the manufacturer´s instruction (BioRad, US).

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Discussion

Discussion

5 years ago

Author: Hollie Fowler United Kingdom

How can I deal with my pellet being too viscous?

I am using 8M Urea to lyse my cells but after centrifugation my pellet is very viscous. This causes me trouble when I try to collect the supernatant. Is there any way to avoid it and do you think it will have an effect on the concentration of my protein?

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Papers

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for B-PER™ Bacterial Protein Extraction Reagent below.

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