This antibody portfolio includes a wide range of tissue- & tumor-specific primary antibodies for basic research, product development and analysis of a variety of biological questions. The monoclonal antibodies are perfectly applicable for all cell or tissue biology studies and for the analysis of predictive markers in research. High epitope affinity Highly specific detection of the target protein Unconjugated Suitable for e.g. western blot, immunoprecipitation, ELISA & immunofluorescence anti-Cyclin D1 mouse monoclonal, DCS-6 100 μg/ml, monoclonal, affinity purified DCS-6 reacts specifically with cyclin D1 protein present predominantly in G1 phase of cell cycle; it does not cross-react with D2 or D3 cyclins (the closest relatives within the cyclin family); DCS-6 is especially useful in tumour diagnosis: it can be used as fast method to identify cases with 11q13 amplifications. Cyclin D1 gene is identical to the BCL-1 and the PRAD-1 oncogene and is part of the chromosome 11q13 amplicon which is amplified in several common tumour types like breast, lung, oesophagus, urinary bladder and head and neck carcinoma. Polypeptide reacting: Mr 36 kDa polypeptide (cyclin D1 polypeptide of human cells). Reactivity on cultured cell lines: MCF-7, MDA-MB-134 and MDA-MB-231 (breast cancer cell lines), UMSC22 (squamous cell carcinoma cell line), U-2-OS (osteosarcoma cell line), RT-112, A431, HT-29, 3T3-L1, Ins-1. The expression of Cyclin D1 is tissue specific and associated with tumour onset and progression. Application Immunocytochemistry (ICC): Assay dependent Immunohistochemistry (IHC) - frozen: 1:20-1:50 (2-5 μg/ml) Immunohistochemistry (IHC) - paraffin: 1:20-1:50 (2-5 μg/ml, microwave treatment recommended) Western Blot (WB): 1:50-1:100 (1-2 μg/ml) IHC analysis of head and neck squamous cell carcinoma using anti-Cyclin D1. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Cyclin D1 mouse monoclonal, DCS-6 (Cat. No. 690053) was diluted in PBS (antibody concentration 2 μg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes. The 20x picture was acquired using microscopy (courtesy of J. Hess, University Hospital Heidelberg). IHC analysis of human liver using anti-Cyclin D1. IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-Cyclin D1 mouse monoclonal, DCS-6 (Cat. No. 690053) was diluted in PBS (antibody concentration 2 μg/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visable and with Haemalaun for a few minutes. The 20x picture was acquired using microscopy (courtesy of J. Hess, University Hospital Heidelberg). Western blot analysis of mouse 3T3-L1 and rat Ins-1 cell lysate with anti-Cyclin D1 antibody. Western blot analysis was performed on 20 μg of either 3T3 or Ins-1 lysate. Cells were lysed with RIPA buffer. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-Cyclin D1 mouse monoclonal, DCS-6 (Cat. No. 690053) was diluted in blocking buffer (antibody concentration 2 μg/ml) and incubated for 1 h at RT. The secondary antibody anti-mouse IgG goat polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 μg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate. Not a medical device / Not an IVD product