Most classical assays are compatible with our LYSE and LYSE-NHS buffers. We recommend the BCA assay or the tryptophan quantification method. Some assays require dilution with destilled water to achieve best results.

Dilutions:
- BCA: none
- Bradford: 1:4
- Coomassie: 1:20
- Lowry: 1:4
- Tryptophan: none

We recommend storage at RT for up to 3 months. The iST and iST-NHS kits are shipped at ambient temperature. Upon arrival, please store the enzyme mix at -20°C and the rest of the kit at room temperature.

We get highly reproducible results with protein starting amounts ranging from 1 µg to 100 µg. For low input information please see “Should I adjust the volumes of the buffers with protein starting amounts?”

Please inquire for further information on the starting amounts for various sample types. A short overview of recommended raw material amounts is also given on the inside of the iST and iST-NHS kits and can be downloaded also on our resources page.

We strongly recommend to use the LYSE or LYSE-NHS buffers provided with the iST or iST-NHS kits, respectively. Sonication (e.g. Diagenode Bioruptor systems) can be used together with the LYSE and LYSE-NHS buffers to increase cell lysis.
If you want to use your own lysis buffer, the efficiency of cell lysis and thus protein identification will likely suffer. Please refer to the following table or contact us to check the compatibility of your lysis buffer with the iST system:

For further information on concentrated LYSE or LYSE-NHS buffers contact us.

For simplicity reasons we recommend usage of buffer volumes indicated in the protocol. You can adjust the volume of LYSE and DIGEST to your starting amounts (e.g.: 20 ug protein starting material, 10 ul LYSE, 10 ul DIGEST, keep the other buffers as indicated in the protocol). For less than 20ug protein starting material, stay with 10ul LYSE and DIGEST, respectively. Accordingly, you may adjust the volumes for chemical labelling and quenching as recommended by the label manufacturer.

We recommend to perform the lysis/denaturation at 95°C and only use lower temperatures for temperature-sensitive samples such as IP samples. We have tested lower temperatures down to 60°C and do not see any differences in parameters tested (IDs, alkylation rates, etc.).

Yes, IP samples can directly be processed with the kit. However, the protocols for Agarose- or magnetic affinity matrices slightly differ, please inquire or refer to our Resources for adapted protocols.

We recommend to perform your standard homogenization in our lysis buffer (e.g. bead milling, grinder, etc.). After homogenization boil the sample (FFPE samples 1h at 95°C) and continue with the protocol as recommended. If you need additional LYSE buffer please contact us.

Yes, FFPE samples can directly be processed with the kit. Please inquire or refer to our website for an adapted protocol.

Yes, urine samples can directly be processed with the kit. However, given the high amount of bilirubin, we provide an extra washing buffer for urine samples. Please inquire or refer to our website for an adapted protocol.

Our LYSE and LYSE-NHS buffers denature, reduce and alkylate proteins. Thus, please consider the following fixed modifications in your database search:
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The iST and iST-NHS Kits 96 rxn allow you to start immediately with ready-to-go reagents and offer the flexibility to process either 96 samples at once or multiple batches of smaller sample numbers.

Chemical labeling experiments require very high peptide labeling efficiencies (>98%) for proper quantification. When struggling with lower labeling efficiencies, please see the following recommendations:

Currently, our kits come with an enzyme mix consisting of LysC and trypsin. We are developing solutions for alternative enzymes.

Yes, we ship our products worldwide from Germany. For our US customers, we drop-ship directly from our warehouse in New Jersey.

The Phoenix peptide cleanup kit is able to remove detergents, sugars, lipids, salts and various polymers. For more details see our Phoenix Application Note.

Our heating shaker is compatible with plate-based shaker systems in the SPSS format, e.g. the Eppendorf ThermoMixer® C.

For the convenience of our customers, we offer several ways of purchasing our products:

- Order by email: order@preomics.com
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- Order by phone: +49-89-2314163-0
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- Order by fax: +49-89-2314163-99
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- Order from our webshop: add products to cart, shipping costs and taxes are added at the checkout. Directly pay with credit card or PayPal.

Yes, we ship our products worldwide from Germany. For our US customers, we drop-ship directly from our warehouse in New Jersey.