Anti mum1 clone mum1p

In all cases, bone marrow biopsy evaluation was performed as a part of the staging workup for MM at iliac spine. In selected patients (cases 16, 17, and 18), typical HU < 0 LBLs underwent to needle biopsy. In detail, 3-μm-thick slides were obtained from formalin-fixed, decalcified paraffin-embedded marrow samples. Following the routine protocol for bone marrow examination, the slices were stained with H&E, PAS, and Giemsa. Marrow fibrosis was assessed by reticulin stain. The immunohistochemical characterization of plasma cells was performed using the following antibodies: anti-CD138 (clone MI15 Dako, Glostrup, Denmark), anti-MUM1 (clone MUM1p, Dako, Glostrup, Denmark), anti-CD56 (clone 504 Leica, Milan, Italy), anti-cyclin D1 (clone EP12, Dako, Glostrup, Denmark), anti-CD20 (clone L26, Dako, Glostrup, Denmark), anti-CD79a (clone 11E3, Leica, Milan, Italy), anti-CD3 (clone LN10, Novocastra, Milan, Italy), and Mib1 (clone Mib1 Dako, Glostrup, Denmark). An anti-CD68 antibody (clone PGM1, Dako, Glostrup, Denmark) was used to assess the presence and distribution of peri-trabecular osteoclasts.

IHC was performed on a total of 130 samples using the DAKO EnVison detection Kit (Dako, Glostrup, Denmark) in accordance with the manufacturer’s instructions. Freshly cut 4-um FFPE sections were subjected to heat-induced antigen retrieval in EDTA buffer (PH = 9.0) for 2–3 min. A panel of primary antibodies was utilized in our study, including anti-CD10 (clone 56C6, 1:50, Novocastra), anti-BCL6 (clone PG-B6p, 1:40, Dako), anti-MUM1 (clone MUM1p, 1:50, Dako), anti-BCL2 (clone 124, 1:100, Dako), anti-C-MYC (clone EP121, 1:75, Zhongshan) and anti-P53 (DO-7, 1:100, Dako). IHC staining was performed with colour development carried out using 3,3′-diaminobenzidine tetrahydrochloride.
According to the requirements and recommendations of the 2016 revised WHO classification system, the expression statuses of BCL2 and C-MYC were identified in all samples. Cutoff values of 50% for BCL-2 expression and 40% for C-MYC expression was used to identify DEL [9 (link)]. Germinal centre B-cell (GCB)/non-GCB types were grouped according to Han’s algorithm [14 (link)], which is based on immunostaining against CD10, BCL-6 and MUM1, using a cut-off of 30% for each antibody.