GentleLys - Native Cell Lysis Buffer

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GentleLys Buffer Type

GentleLys Dissolve

GentleLys Stabilize

Volume

Quantity

Description

The Cube Biotech GentleLys Buffer is the only 100% detergent-free, non-denaturing solution to lyse cultured cells from plated cells as well as cells pelleted from suspension cultures while solubilizing all membranes in the process. GentleLys enables the extraction of soluble proteins and the stabilization of membrane proteins from all compartments of the cells. It is compatible with many applications, including Immunoassays, Protein Assays, Mass Spectrometry, and Protein Purification.
The Buffer comes in two variants: GentleLys – Dissolve which lyses cells in 15 min while GentleLys – Stabilize lyses all cells and makes the whole proteome (soluble and membrane proteins) of the cell available for your analytic approaches. It solubilizes all membranes and stabilizes expressed membrane proteins in their native conformation
They are able to do this thanks to the inclusion of synthetic nanodisc copolymers. More on that can be found here.

Note: GentleLys buffers only work on eucaryotic cells. Bacterial cells cannot be lyzed using GentleLys buffers. Another cell type to keep in mind are plant cells. It is NOT possible to treat intact plant cells with GentleLys due to the cell wall. But treatment of the protoplast (cell wall removed) is possible!

Datasheets

Lab Results

Overview of the capabilities of both GentleLys Buffer Types — **Figure 1:** Overview of both types of the GentleLys types. They can both be used for almost the same applications. The only difference is that GentleLys Dissolve cannot stabilize membrane proteins, while GentleLys Stabilize is able to do that.

Amount of detected living cells GentleLys vs RIPA — **Figure 2:** Total number of countable cells after 1 min / 10 min treatment with either RIPA buffer, GentleLys – Dissolve / GentleLys - Stabilize or control buffer. Lower cell count equals fully lysed cells. The less cells are countable the more sufficient the cells lysis takes place (no organelles detectable). Note that there is no graph for GentleLys - Stabilize after 1 minute of treatment. This is because we recommend this treatment for at least 15 minutes.

Comparison RIPA buffer vs GentleLys Dissolve Buffer using HEK293 cells - SDS Page; Quantification of comparison between RIPA buffer and GentleLYs buffer in HEK293 cells — **Figure 3: A** SDS PAGE analysis of HEK293 cell supernatant after 1 min / 10 min / 30 min treatment with either RIPA buffer, GentleLys – Dissolve or control buffer. After incubation intact cells and cell debris were centrifuged and equal volumes of supernatant were run via SDS PAGE and detected via Coomassie staining. B Quantification of accumulated data shown in A. Intensity of individual band patterns have been measured. With GentleLys a ~20% increased protein amount can be identified via SDS PAGE band analysis. RIPA has been set to 100%.

Comparison RIPA buffer vs GentleLys Dissolve Buffer using T.ni cells - SDS Page; Quantification of comparison between RIPA buffer and GentleLYs buffer in T.ni cells — **Figure 4 A:** SDS PAGE analysis of T.ni. cell supernatant after 5 min / 10 min tretment with either RIPA buffer, GentleLys – Dissolve or control buffer. After incubation intact cells and cell debris were centrifuged and equal volumes of supernatant were run via SDS PAGE and detected via Coomassie staining. B Quantification of accumulated data shown in A. Intensity of individual band patterns have been measured. With GentleLys a ~50% increased protein amount can be identified via SDS PAGE band analysis. RIPA has been set to 100%.

FAQ

Can I get the datasheets for the GentleLys Buffers?

Of course, you can find them here:

Why should I use this buffer instead of the RIPA buffer?

There are two major reasons for that:

The GentleLys buffer keep the native folding of your proteins intact. (Hence the name "Gentle"). The RIPA buffer is denaturing and does not do that.
The RIPA buffer uses detergents, while the GentleLys buffer uses copolymers. Detergents do not allow for protein-ELISAs as a follow-up assay. Copolymers do not have this limitation.

What other downstream assay can samples created through the GentleLys buffer be used for?

There are numerous applications for samples obtained through the use of the GentleLys buffers.

Protein-ELISA

What are the storage conditions for the GentleLys Buffers?

The Buffers can be stored at 2-8°C.

What cell types can the GentleLys Buffer be used for?

The buffer is primarily meant to be used for insect cells and mammalian cells. Bacterial cells and plant cells do not work due to the cell wall.

What is the shelf-life of GentleLys buffers?

The lyophilized 1ml sample can be stored for 2-3 years if the bottle remains unopened.

For the 100 ml and 250 ml bottles of liquid buffer, the product can be stored for up to 6 months after delivery.

Can I use the Stabilize Buffer also to simply dissolve my cells like the Dissolve Buffer does?

Both Buffer variants are capable of dissolving the cells. However, only the Stabilize Buffer is capable of stabilizing the membrane proteins.

How does the membrane protein stabilization of the GentleLys Buffer work?

Through the use of our specially engineered copolymer.More on that can be found here:

Disclaimer

Our products are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.