| Immunogen | Clinical Tissue Human Breast Normal Tissue Lysate |
| Specificity | Tissue: Breast Diagnosis:Infiltrating ductal carcinoma Stage:IIIB TNM:T4bN1M0 Grade:2 Sex: Female Age :54 Gross findings: Tumor size 10 cm., ill demarcated. Ulceration of the skin. Cut section firm and white. Ipsilateral axillary lymph nodes examined: 2 Microscopic findings: The tumor is composed of proliferation of malignant epithelial cells. The epithelial cells are arranged as groups which have invaded the stroma. The tumor cells have irregular , basophilic nuclei and abundant cytoplasm. Nuclear chromatin is coarse with prominent nuclei. Mitotic figures are evident. The stroma is infiltrated by small numbers of chronic inflammatory cells. Necroses are seen but blood vessel invasions are not revealed. The lymph nodes show metastasis (2/8). |
| Preparation Method |
Tissue specimens are homogenized in modified RIPA buffer to obtain the soluble proteins, and centrifuged to clarify. The pellet was further extracted with a second buffer to obtain the less soluble protein fraction. The lysate solution may appear turbid at cold temperatures due to insolubility of buffer components. The solution should clear upon warming to room temperature. |
| Application Notes | These lysates have not been subjected to denaturing or reducing conditions. This allows the tissue or cell lysate to be used in a variety of applications; to study protein-protein interaction, ligand binding, ELISA, immunoprecipitation, 1D and 2D gel electrophoresis, and Western blotting for the detection of specific protein targets. For use in 1D and 2D gel electrophoresis, the addition of a denaturing gel loading buffer with reducing agents may be required. Buffer requirements for performing protein-protein interaction and ligand binding studies can vary significantly from RIPA buffer and may require modifications. In most cases, tissue lysates in RIPA buffer can be used, directly in standard ELISA and immunoprecipitation assays. This material has tested negative for HbsAg, HIV 1/2, and HCV. Use UNIVERSAL PRECAUTIONS when handling. Human tissue derivatives must be treated as a potentially infectious agent and disposed of appropriately. |
| Storage | Store at -80C. Avoid freeze-thaw cycles. |
| Concentration | 1 mg/ml |
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