FAQ

Frequently asked questions

We recommend storage at RT for up to 6 months. The kit is shipped at ambient temperature but please store the enzyme mix at -20°C.
We get highly reproducible results with protein starting amounts ranging from 1 µg to 100 µg.
Please inquire or refer to our website 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 kit.
We strongly recommend the LYSE buffer provided with the iST kit, which was extensively optimized in order to achieve best results. Sonication (e.g. Diagenode Bioruptor systems) can be used together with the LYSE buffer to increase cell lysis. If you want to use your own lysis buffer, the efficiency of cell lysis and thus protein identification might be reduced. Please contact us to check the compatibility of your lysis buffer with the iST system. Protein precipitation might be necessary in certain cases.
Yes, IP samples can directly be processed with the kit. Please inquire or refer to our website for an adapted protocol.
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.
Most classical assays are compatible with our lysis buffer. We recommend the BCA assay or the Tryptophane quantification method. Some assays require dilution with water to achieve best results.

Dilutions:

  • BCA: none
  • Bradford: 1:4
  • Coomassie: 1:20
  • Lowry: 1:4
  • Tryptophan: none
The LYSE buffer reduces and alkylates proteins. Thus, carbamidomethylation on cysteine residues has to be set as a fixed modification: (C2H3NO), +57 Da mass shift, Unimod modification #4.
We provide aliquots of all critical reagents to guarantee best quality and that you can perform the whole sample preparation with lower sample numbers.
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.
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.

Any more questions?
Don’t hesitate, contact us!