Anti human mitochondria antibody

The use of human cells and animals were approved by the NUS Institutional Review Board, Institutional Biosafety Committee and Institutional Animal Care and Use Committee (R17-0956, 12/12/2017).
Human dental pulp stem cells (DP003F, Alcells, Alameda, USA) were cultured in basal growth culture medium ((Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Carlsbad, CA, USA), 10% fetal bovine serum (Invitrogen) and 1% penicillin/streptomycin (Invitrogen)) until 70~80% confluence and harvested (TrypLE, Invitrogen). As shown in Figure 7B, MSC (2 × 10⁴, passage 3) were seeded in the graphene scaffold (0.8 × 0.8 × 0.2 cm) and maintained undisturbed for seven days. Following that, the scaffolds (n = 6) were placed inside 3D-printed polylactide protection containers (Cube, 3D Systems, Columbia, SC, USA) and transplanted into the subcutaneous space of immunodeficient mice (CB-17 SCID, Invivos, Singapore). After 28 days, specimens were retrieved, fixed with 4% formaldehyde solution in phosphate-buffered saline at 4 °C for 24 h and stained with hematoxylin and eosin (H&E). Immunohistochemical analyses of the tissues formed were performed for OPN and RUNX2 (1:1000, Abcam, Cambridge, UK), and anti-human mitochondria antibodies (1:500, Abcam). Controls were tissue sections stained with an isotype-matched non-specific IgG antibody.

A section of each tumor was fixed in 4% paraformaldehyde blocks and the morphology was compared to that of the original patient tumor by H&E staining and IHC analysis. The following primary antibodies were used: CGA (1:100; Abcam, Cambridge, UK), SYP (1:50; Santa Cruz), rabbit anti-human anti-PSA, anti-AR monoclonal antibodies (1:100, Abcam, Cambridge, UK). Anti-AR(AR-V7 specific) antibodies (1:200, Abcam, Cambridge, UK) and anti-human mitochondria antibodies (1:100, Abcam, Cambridge, UK). The secondary antibody was a biotin-labeled goat anti-rabbit, which was used according to a published protocol [1 (link)].