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. Author manuscript; available in PMC: 2022 Mar 10.
Published in final edited form as: J Proteome Res. 2020 Jun 8;19(8):2989–2996. doi: 10.1021/acs.jproteome.0c00024

Figure 3.

Figure 3.

Generalized workflow of Endo F3 and PNGase F treatments on a tissue. Begin by cutting the tissue from the FFPE block at 5 μm onto the slide; (I) heat the slide at 60 °C for 1 h; (II) dewax in the series of xylene, ethanol dilutions, and water, antigen retrieval in citraconic buffer; (III) apply Endo F3 to the tissue; (IV) incubate in a humidity chamber at 37 °C for 2 h; (V) apply the CHCA matrix to the tissue; (VI) Image on MALDI-FT-ICR; (VII) Clear the matrix and glycans with ethanol dilutions, high-pH and low-pH washes; (VII) apply PNGase F to the tissue; (IX) incubate in the humidity chamber at 37 °C for 2 h; (X) Apply the CHCA matrix to the tissue; (XI) Image on MALDI-FT-ICR.