6-异硫氰酸荧光素

产品简介

外观：黄色粉末

纯度：≥95% (HPLC)

产品参数

Ex(nm)：494

Em(nm)：520

Solvent：DMSO

产品描述

FITC能和各种抗体蛋白结合，结合后的抗体不丧失与一定抗原结合的特异性，并在碱性溶液中具有强烈的黄绿色荧光。通过在荧光显微镜下观察或流式细胞仪分析可对相应抗原进行定性、定位或定量的检测。用于医学，农学和畜牧等方面，可对由地细菌病毒和寄生虫等所致疾病进行快速诊断。

FITC标记抗体流程

（1）将待交联的蛋白（浓度≥1mg/mL）对交联反应液透析三次4℃，至pH＝9.0。交联反应液配制方法：7.56g NaHCO3，1.06g Na2CO3，7.36g NaCl，加水定容至1 L。

（2）将FITC溶于DMSO中，浓度为1mg/mL。每次交联使用的FITC均应新鲜配制，避光。

（3）按P:F（蛋白质:FITC）=1mg:150μg 的比例将FITC缓慢加入于抗体溶液中，边加边轻轻晃动使其与抗体混合均匀，暗处4℃反应8h。

（4）加入5mol/L的NH4Cl至终浓度50mmol/L，4℃终止反应2h。

（5）将交联物在PBS中透析四次以上，至透析液清亮。

（6）交联物的鉴定

蛋白浓度(mg/mL) = [ A280– 0.31×A495 ] / 1.4

F/P比例: 3.1×A495 / [A280 – 0.31×A495 ]，该值应介于2.5～6.5之间。

（7）FITC交联的蛋白应置于pH 7.4的磷酸盐缓冲液中，加入0.1% NaN3、1% BSA，4℃避光保存。

Product Introduction

Appearance: yellow powder

Purity: ≥95% (HPLC)

Product parameter

Ex(nm): 494

Em(nm): 520

Solvent: DMSO

Product description

FITC can bind to various antibody proteins, and the bound antibody does not lose the specificity of binding to a certain antigen, and has strong yellow-green fluorescence in alkaline solution. The corresponding antigen can be qualitatively, localized or quantitatively detected by observation under a fluorescence microscope or flow cytometry analysis. Used in medicine, agriculture and animal husbandry, it can quickly diagnose diseases caused by bacteria, viruses and parasites.

FITC labeled antibody process

(1) The protein to be crosslinked (concentration ≥1mg/mL) was dialyzed three times against the crosslinking reaction solution at 4°C to pH=9.0. The preparation method of the crosslinking reaction solution: 7.56g NaHCO3, 1.06g Na2CO3, 7.36g NaCl, add water to make the volume to 1 L.

(2) Dissolve FITC in DMSO at a concentration of 1 mg/mL. FITC used for each cross-linking should be freshly prepared and protected from light.

(3) Slowly add FITC to the antibody solution at the ratio of P:F (Protein: FITC) = 1mg: 150μg, and gently shake to mix it with the antibody while adding, and react at 4°C in the dark for 8 hours.

(4) Add 5mol/L NH4Cl to a final concentration of 50mmol/L, and terminate the reaction at 4°C for 2h.

(5) Dialyze the cross-linked substance in PBS for more than four times until the dialysate is clear.

(6) Identification of cross-linked products

Protein concentration (mg/mL) = [A280– 0.31×A495] / 1.4

F/P ratio: 3.1×A495/ [A280 – 0.31×A495], the value should be between 2.5 and 6.5.

(7) FITC cross-linked protein should be placed in a pH 7.4 phosphate buffer, added with 0.1% NaN3, 1% BSA, and stored at 4°C in the dark.

Application：

6-Fluorescein Isothiocyanate is a fluorescent reagent used in fluorescence polarization binding assay to develop inhibitors of inactive p38α Mitogen-activated protein kinase (MAPK).