Cyclin D1

Cyclins are a family of proteins involved in the progression of cells through the cell cycle. Cyclins form a complex with their partner, cyclin-dependent kinase (Cdk), which activates the latter’s protein kinase function. Cyclins are so named because they are produced or degraded as needed in order to drive the cell through the different stages of the cell cycle. When its concentrations in the cell are low, the cyclin detaches from the Cdk, inhibiting the enzyme’s activity, probably by causing a protein chain to block the enzymatic site.

Cyclin D1 or PRAD-1 or bcl-1 is one of the key cell-cycle regulators, and functions in association with Cdk4 and/or Cdk6 by phosphorylating the Rb protein. It is a putative proto-oncogene overexpressed in a wide variety of human neoplasms including Mantle Cell Lymphomas (MCL).

Cyclin D1 is a rabbit 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.

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1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. 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).
5. 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.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.