Learn! Access our knowledge bank

The protocol specifies a temperature range of 80-95 °C and temperatures at the upper end should increase extraction and de-crosslinking efficiency.

If you use cap strips to seal the BeatBox 96w PLATE and a thermoshaker for heating, please note the following: The temperature reached inside the wells may vary between different thermoshaker models. At very high temperatures the cap strip may burst open due to high vapor pressure of the lysis buffer. To avoid sample loss, perform a test run with lysis buffer to identify the highest possible temperature for your thermoshaker setup. We do not recommend using a heated lid (e.g. ThermoTop # 5308000003 Eppendorf). Use a conventional lid instead. If only a ThermoTop is available, reduce the temperature to 80-85°C.

A lower heating temperature may sometimes lead to lower protein extraction efficiency. To improve results, you can perform a second cycle of BeatBox and heating.

Alternatively, use a PCR cycler with a heated lid exerting pressure from the top and close the BeatBox 96w PLATE with the SILICONE MAT. This allows you to select a higher temperature for the de-crosslinking step.

Depending on the amount of input material, you may have to perform multiple rounds of BeatBox and heating to help with homogenization/hydration of the tissue. We recommend testing the following sequence: 10 min BeatBox -> 30 min at 80-95 °C -> 10 min BeatBox -> 30 min at 80-95 °C. If the tissue is still not fully homogenized, continue with another cycle of BeatBox and heating.

Yes, this is possible. FFPE tissue homogenates in iST LYSE or SP3 LYSE can be frozen to be processed at a later timepoint. Just note that if you homogenized paraffin containing FFPE samples, the paraffin dispersed in the solution will precipitate. After thawing the sample, please heat to >60°C until the paraffin is dissolved.

NOTE: For freezing, please follow the instructions for use for Eppendorf Safe-Lock Tubes and do not use the tubes with liquid nitrogen.

We currently do not recommend using the BeatBox Tissue Kit 24x.

Yes, this is possible. We recommend replacing the acidic SP3 LYSE supplied with the kit with a different lysis buffer with a higher pH, e.g. 50 mM TEAB (pH 8.5) with 5% SDS or RIPA buffer. Lysis buffers with basic pH typically show better extraction performance with FFPE tissue.For a modified protocol, please contact us at info@preomics.com

Yes, these buffers can also be used for the protein extraction step. If the SDS concentration in the sample is 0.1%, the iST workflow remains unchanged. For higher amounts of SDS, use the SP3-iST Add-on kit to remove SDS before continuing with the DIGEST step of the iST workflow. After protein clean-up on the SP3-beads, residual paraffin can still be still present in the sample. We recommend using WASH 0 for full paraffin removal during iST clean-up, as outlined in the FFPE protocol.

The Metal Heating Shaker Adapter guarantees optimal heat transfer for our CARTRIDGES compared to planar heating systems. It is compatible with any heating shaker in the SPSS format and many liquid handling platforms. It is also directly compatible with our 96-well CARTRIDGE adapter plates.

All our kits in the 96 reaction format already contain the 96-well adapter plate required for convenient handling of larger sample numbers.

Customers from North America please order via PreOmics Inc. (USA), customers from the rest of the world please order via PreOmics GmbH (Germany)

We offer several options to order our products:

• Send an email to: order@preomics.com‍
• Call us: +49-89-2314-163-0‍
• Fax us: +49-89-2314-163-99

‍

Yes, we do ship our products worldwide. For the convenience of our US customers, we ship from our warehouse in New Jersey.

We provide complete solutions to ensure best results for your LC-MS/MS analyses. Adaptions to our lysis buffers might be necessary though for specific experimental questions. Upon request, we provide the appropriate 2x concentrated LYSE buffer and the WASH0 for use with plant, algae and diatom samples.

We ship at ambient temperature. Upon arrival, please store the lyophilized enzyme mix (red DIGESTtubes) at -20°C and the rest of the kit at room temperature.

No, freezing is detrimental to our buffers. Only the lyophilized enzymes (red DIGEST tubes) in the iST, iST-REG-PSI, iST-NHS and iST-BCT kits should be frozen upon arrival for long-term storage.

Resuspended DIGEST can be stored at 4°C for up to two weeks. For long-term storage, lyophilize the DIGEST again, lyophilized DIGEST can be stored at -20°C for up to nine months.

We guarantee a minimum remaining shelf life of three months after receiving our products. Please refer to the shelf life information printed on each kit box for further details.

The performance of our LYSE / LYSE-NHS / LYSE-BCT will drop significantly after the expiration date. Thus, we do not recommend to use our kits after the shelf life has expired.