儲 存 液: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol
保存條件: Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
Anti-E cadherin抗體,上皮鈣粘附分子抗體產品介紹:Cadherins comprise a family of Ca2+-dependent adhesion molecules that function to mediate cell-cell binding critical to the maintenance of tissue structure and morphogenesis. Members of this family of adhesion proteins include rat cadherin K (and its human homolog, cadherin-6), R-cadherin, B-cadherin, E/P cadherin and cadherin-5. The classical cadherins, E-, N- and P-cadherin, consist of large extracellular domains characterized by a series of five homologous NH2 terminal repeats. The most distal of these cadherins is thought to be responsible for binding specificity, transmembrane domains and carboxy terminal intracellular domains. The relatively short intracellular domains interact with a variety of cytoplasmic proteins, such as ∫-catenin, to regulate cadherin function.
免疫組化問題解答:
1 染色過強
原因 解決方法 a 抗體濃度過高或孵育時間過長 降低抗體滴度、抗體孵育時間:室溫1小時或4度過夜 b 孵育溫度過高超過37度 一般在室溫20-28度 c DAB顯色時間過長或濃度過高 顯色時間不超過5-12分鐘,以顯微鏡下觀察為準 2 非特異性背景染色
原因 解決方法 a 操作過程中沖洗不充分 每步沖洗3次每次5分鐘 b 組織中含過氧化物酶未阻斷 可再配置新鮮3%H2O2封閉孵育時間延長 c 組織中含內原性* 正常非免疫動物血清再封閉 d 血清蛋白封閉不充分 延長血清蛋白封閉時間
3 染色弱
原因 解決方法 a 抗體濃度過低、孵育時間過短 提高抗體濃度、孵育時間不能少 于60分鐘 b 試劑超過有效使用時間 應更換試劑 c 操作中添加試劑時緩沖液未瀝干 每步滴加試劑前瀝干切片中多余的緩沖液 使試劑稀釋 (應防止切片干燥) d 室溫太低 低于15度 要改放在37度孵育箱孵育30-60分鐘或4 度冰箱過夜 e 蛋白封閉過度 封閉不要超過12分鐘
4 染色陰性
原因 解決方法 a 操作步驟錯誤 應重試 設陽性對照 b 組織中無抗原 設陽性對照以驗證試驗結果 c 一抗與二抗種屬連接錯誤 仔細確定一抗二抗種屬無誤。