異硫氰酸熒光素FITC是生化試劑,也是醫學診斷藥物,主要用于熒光抗體技術中的熒光染料,能和各種抗體蛋白結合,結合后的抗體不喪失與一定抗原結合的特異性,并在堿性溶液中具有強烈的綠色熒光。用于醫學,農學和畜牧等方面,可對由地細菌病毒和寄生蟲等所致疾病進行快速診斷。
異硫氰酸熒光素標記用試劑,多肽、蛋白、抗體標記用試劑,氨基標記用熒光試劑,含異硫氰基的熒光試劑。
艾美捷異硫氰酸熒光素相關參數:
中文名稱:異硫氰酸熒光素
英文名字:FITC [FITC Isomer I; fluorescein-5-isothiocyanate]
貨號:B-CHM100-100mg
規格:100mg
產品形式:固體
保存建議:異硫氰酸熒光素儲存條件:-20℃干燥避光保存,有效期一年。
純度:高于或等于95%(通過高效液相色譜法測定)
光譜特性:激發/發射波長 = 490 / 525 納米
外觀:黃色粉末
異硫氰酸熒光素標記抗體流程:
(1) Dialyze the protein to be cross-linked (concentration: 1 mg/mL) against the cross-linking reaction buffer threetimesat 4°C, adjusting the pH to 9.0. Preparation of the cross-linking buffer: Dissolve 7.56 g NaHCO3, 1.06 g Na2CO3, and7.36g NaCl in water to a final volume of 1 L.
(2) Dissolve FITC in DMSO at a concentration of 1 mg/mL. Prepare fresh FITC solution for each cross-linking reactionandavoid exposure to light.
(3) Add FITC slowly to the antibody solution at a ratio of P:F (protein:FITC) = 1 mg:150 μg. Gently mix the solutionwhileadding FITC to ensure uniform mixing with the antibody. Allow the reaction to proceed at 4°C in the dark for 8hours.
(4) Add 5 mol/L NH4Cl to a final concentration of 50 mmol/L to stop the reaction at 4°C for 2 hours.
(5) Dialyze the conjugates against PBS solution at least four times until the dialysate becomes clear.
(6) Characterization of the cross-linked material:Protein concentration (mg/mL) = [A280 - 0.31 × A495] / 1.4F/P ratio: 3.1 × A495 / [A280 - 0.31 × A495]. This value should be between 2.5 ~ 6.5.
(7) Store the FITC protein conjugates in a PBS at pH 7.4, supplemented with 0.1% NaN3 and 1%BSA, storeat 4°Cinthe dark
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