Anti ki 67 mib 1 antibody

Surgical or biopsy specimens were fixed in 10% formalin and were embedded in paraffin. The histological tumor type was established based on specimens stained with hematoxylin and eosin, according to the previous WHO criteria [22 (link)]. The relevance of this investigation is limited by the fact that the tumor classification is based on the previous, out-of-date WHO classification and so may no longer be directly applicable to the current classification [24 (link)].
The cellular proliferation activity of the tumor was determined by measuring the MIB-1 proliferation index obtained by immunohistochemical staining with anti-Ki-67/MIB-1 antibody (Dako, Tokyo, Japan). The percentage of tumor cells stained positively for MIB-1 antigen (MIB-1 labeling index: MIB-1 LI) was determined in the area containing the largest number of positive tumor cells and was regarded as representative of the tumor proliferation activity.

Tissue sections were stained with an anti-ATRX polyclonal antibody (1: 300 dilution, ATLAS Antibodies AB, Bromma, Sweden), anti-GFAP (6F2) monoclonal antibody (1: 200 dilution, DAKO, Glostrup, Denmark), anti-IDH1 R132H (H09) monoclonal antibody (1:300 dilution, Dianova, Hamburg, Germany), anti-Ki67 (MIB-1) antibody (1:100 dilution, DAKO), anti-H3K27 M polyclonal antibody (1:700 Millipore, Temecula, USA), anti-p53 monoclonal antibody, DO-7 code M7001 (1:100 dilution, DAKO), anti-PHH3 antibody (1:100 dilution, Cell Marque, Rocklin, CA, USA) and anti-vimentin antibody (1:500, DAKO) (Supplementary Table 2). IHC staining was performed using a standard avidin–biotin-peroxidase method with a BenchMark ULTRA system (Roche Diagnostics, Indianapolis, US). The positive control was known positive tissue, and entrapped positive cells were used. For the negative control, the primary antibody was omitted.
ATRX gene mutations were scattered throughout exons and introns, making some sites difficult to capture using NGS. Since the initial version of our customized brain tumor panel could not detect full variants of ATRX mutations, we relied on ATRX IHC, but upgrade version of BTP panel could detect all the mutations.