Tubulin-Tracker Green

Tubulin-Tracker Green是一种Tubulin绿色荧光探针，可以用于培养细胞或组织切片的Tubulin特异性荧光染色。本Tubulin-Tracker Green探针为荧光染料Alexa Fluor 488标记的抗α-Tubulin小鼠单克隆抗体(克隆号为DM1A)，可以识别α-Tubulin的426-450aa。可以用于人、小鼠、大鼠、牛、猪、豚鼠和禽类(avian)样品α-Tubulin的检测。最大激发波长为495nm，最大发射波长为519nm。Alexa Fluor 488的荧光光谱和FITC非常接近，可以用FITC的检测条件进行检测。本产品可以用于细胞或组织内的微管(microtubule)的荧光检测。可以用于荧光显微镜观察，也可用于流式细胞仪检测。使用本产品染色细胞内微管的效果请参考图1。图1.Tubulin-Tracker Green对NRK-52E细胞(大鼠肾小管上皮细胞)的染色效果。图A为明场下大鼠肾小管上皮细胞形态图；红色荧光为Mito-Tracker Red CMXRos 染色的线粒体(图B)，绿色荧光为在Mito-Tracker Red CMXRos染色后经固定、通透再使用Tubulin-Tracker Green (抗体法微管绿色荧光探针) 对微管进行的染色(图C)，图D为Mito-Tracker Red CMXRos、Tubulin-Tracker Green及Hoechst 33342三重染色的叠加图。实测数据会因实验条件、检测仪器等的不同而存在差异，图中数据仅供参考。本产品使用时的推荐稀释比例为1:50-100，共可以配制2-4ml染色液。如果每个片子需要使用100µl染色工作液，每个包装的本产品足够用于20-40个片子的染色。-20℃避光保存，1年有效。对于微量的液体，每次使用前先离心数秒钟，使液体充分沉降到管底。荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭。需自备盖玻片和载玻片(可以向阿拉丁订购)。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明 ： 1.固定细胞或组织切片的荧光染色： a.用PBS洗涤细胞或组织切片2次。 b.用阿拉丁的免疫染色固定液或PBS配制的3.7%甲醛溶液室温固定细胞或组织切片约10-20分钟。 c.用阿拉丁的免疫染色洗涤液或含0.1% Triton X-100的PBS洗涤2-3次，每次5分钟。 d.用阿拉丁的免疫荧光染色二抗稀释液或含有1-5%BSA和0.1% Triton X-100的PBS按照1:50-100的比例稀释Tubulin-Tracker Green，例如4µl Tubulin-Tracker Green用0.2ml或0.4ml稀释液稀释，稀释后的溶液即为Tubulin-Tracker Green染色工作液。稀释比例可以根据实际染色效果进行适当调整。 e.把Tubulin-tracker Green染色工作液按照约每个片子100µl的比例滴加到片子上，室温避光孵育30-60分钟。为防止蒸发，孵育时最好将片子置于载玻片染色盒中，载玻片染色盒可以向阿拉丁订购。 f.用阿拉丁的免疫染色洗涤液或含0.1% Triton X-100的PBS洗涤2-4次，每次5分钟。 g.随后可以直接用荧光显微镜进行观察。也可以用抗荧光淬灭封片液封片后保存和观察。 2.流式细胞仪检测： a.每个样品收集约20-50万细胞。 b.PBS洗涤一次。 c.用阿拉丁的免疫染色固定液或PBS配制的3.7%甲醛溶液室温固定细胞约10-20分钟。 d.用阿拉丁的免疫染色洗涤液或含0.1% Triton X-100的PBS洗涤2-3次，每次5分钟。 e.用阿拉丁的免疫荧光染色二抗稀释液或含有1-5%BSA和0.1% Triton X-100的PBS重悬细胞，然后按照1:50-100的比例加入Tubulin-Tracker Green。例如1µl Tubulin-Tracker Green加入到50-100µl细胞悬液中。Tubulin-Tracker Green稀释比例可以根据实际染色效果进行适当调整。 f.室温避光孵育1小时。 g.用阿拉丁的免疫染色洗涤液或含0.1% Triton X-100的PBS洗涤2-3次，每次5分钟。随后即可进行流式细胞仪分析。 For trace amounts of liquid, centrifuge briefly before each use to collect the liquid at the bottom of the tube.Fluorescence intensity decays. Avoid light to minimize the quenching of fluorescence.Coverslips and slides are required but not supplied. They can be ordered from .This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use ： 1. Staining of fixed cells or tissue sectionsa. Wash cells or tissue sections twice with PBS.b. Fix cells or tissue sections at room temperature for 10-20 minutes with 's Immunol Staining Fix Solution or PBS containing 3.7% formaldehyde.c. Wash cells or tissue sections 2-3 times for 5 minutes each with 's Immunol Staining Wash Buffer or PBS containing 0.1% Triton X-100.d. Dilute Tubulin-Tracker Green in a 1:50-100 ratio with 's Immunol Fluorescence Staining Secondary Antibody Dilution Buffer or PBS containing 1-5% BSA and 0.1% Triton X-100. For example, dilute 4µl of Tubulin-Tracker Green in 0.2ml or 0.4ml of dilution buffer to obtain the Tubulin-Tracker Green working solution. The dilution ratio can be adjusted according to the staining result.e. Add 100µl of Tubulin-Tracker Green working solution onto each slide and incubate for 30-60 minutes at room temperature in the dark. To minimize evaporation, it is better to place the slide in a Staining Box for Microscope Slides (, FSR958).f. Wash the slides 2-4 times for 5 min each with 's Immunol Staining Wash Buffer or PBS containing 0.1% Triton X-100.g. The stained slide can be directly observed with a fluorescence microscope. The slides can also be observed or stored after mounting with anti-fade mounting medium.2. Staining of cells for flow cytometric analysis a. Collect approximately 0.2-0.5×106 cells per sample.b. Wash once with PBS.c. Fix the cells at room temperature for 10-20 minutes with 's Immunol Staining Fix Solution or PBS containing 3.7% formaldehyde.d. Wash the cells 2-3 times for 5 minutes each time with 's Immunol Staining Wash Buffer or PBS containing 0.1% Triton X-100.e. Resuspend cells with 's Immunol Fluorescence Staining Secondary Antibody Dilution Buffer or PBS containing 1-5% BSA and 0.1% Triton X-100, then apply Tubulin-Tracker Green at a ratio of 1:50-100. For example, add 1µl of Tubulin-Tracker Green to 50-100µl of cell resuspension. The dilution ratio of Tubulin-Tracker Green can be adjusted according to the staining results.f. Incubate for 1 hour at room temperature in the dark.g. Wash 2-3 times for 5 minutes each with 's Immunol Staining Wash Buffer or PBS containing 0.1% Triton X-100. h. Examine cells by Flow cytometry.

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