Pkm2 antibody

The target cells were cultured in six-well plates until the density reached 50–60%. Next, the cells were fixed using paraformaldehyde, and incubated with an immunofluorescent blocking solution for 1 h. Then, the cells were dehydrated with a gradient alcohol series, soaked in wax, and embedded in paraffin. The sections were boiled in retrieval solution for 10–15 min and allowed to cool to room temperature.
The probe for CTSLP8 was designed to be Cy3 modified, and it was synthesized by Servicebio. The detailed sequence is as follows: 5'-Cy3-GGTTTTAACCTGATCCTTCACAGGACTCAT-3'. First, the probe for CTSLP8 was detected by matched FISH kit in Servicebio. The sections were then incubated with c-Myc antibody (Proteintech; 10,828–1-AP) and PKM2 antibody (Proteintech; 15,822–1-AP). Next, the secondary antibodies of different species were added, and the sections were incubated for 50 min. Finally, the images were captured by confocal microscope (Olympus; FV1000).

Western blotting was performed in accordance with the standard protocols. In brief, cellular proteins were extracted using the RIPA lysis buffer, separated on SDS-PAGE gels, and transferred onto NC membranes (Millipore, Billerica, USA). After incubated in primary and second antibodies separately, proteins were detected by ECL reagent (Millipore, USA) and imaged with FluorChem Q (Protein simple, USA). The following antibodies were used for western blotting: PKM2 antibody (Proteintech, China, 1 : 1000, 15822-1-AP), vimentin antibody (Proteintech, China, 1 : 1000, 10366-1-AP), E-cadherin antibody (Proteintech, China, 1 : 1000, 20874-1-AP), N-cadherin antibody (Proteintech, China, 1 : 1000, 22018-1-AP), and GAPDH antibody (ZSGB-bio, China, 1 : 1000, TA-8).