
bcl-6 is a transcriptional regulator gene which codes for a 706-amino-acid nuclear zinc finger protein. Antibodies to this protein stain the germinal center cells in lymphoid follicles, follicular cells and interfollicular cells in Follicular Lymphoma, Diffuse Large B-Cell Lymphomas, Burkitt’s Lymphoma, and the majority of the Reed-Sternberg cells in Nodular Lymphocyte-Predominant Hodgkin’s Disease.
bcl-6 is also useful in identifying neoplastic cells in cases of nodular Lymphocyte-Predominant Hodgkin’s Disease. In contrast, anti-bcl-6 rarely stains Mantle-Cell Lymphoma and MALT Lymphoma. bcl-6 expression is seen in approximately 45% of CD30+ Anaplastic Large-Cell Lymphomas but is consistently absent in other peripheral T-cell Lymphomas.
For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).
bcl-6 is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.
- Dogan A, Badgi E, et al. Am J Surg Pathol. 2000;24(6):846-852
- Shaffer AL, et al. Immunity. 2000;Vol.13.199-212,Aug.
- M.D. Kraus J, Haley, AM J Surg Pathol. 2000;24(8):1068-78
- Carbone A, et al. Blood. Vol.90,No.6(Sept.15) 1997;pp2445-2450
Recommended Immunohistochemical Protocol
- Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
- Air dry for 2 hours at 58° C.
- Deparaffinize, dehydrate and rehydrate tissues.
- Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
- Any of three heating methods may be used:
- Electric Pressure Cooker
Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature. - Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes. - Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
- Electric Pressure Cooker
- After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
- Wash slides with IHC wash buffer or DI water.
- Continue IHC staining protocol.
Immunohistochemical Protocol
ImmunoDetector
(AP or HRP)
(AP or HRP)