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免疫染色通透液(Saponin)

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库存信息

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货号 (SKU) 包装规格 是否现货 价格 数量
I743382-100ml
100ml 期货 Stock Image
I743382-500ml
500ml 期货 Stock Image
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免疫染色 (36)

基本描述

稳定性与储存 4℃保存,一年有效。-20℃可以长期保存,室温保存一个月内有效。
英文名称 Immunostaining Permeabilization Buffer with Saponin
储存温度 2-8°C储存
运输条件 冰袋运输
产品介绍

产品介绍

免疫染色通透液(Saponin) (Immunostaining Permeabilization Buffer with Saponin, 也称ImmunostainingPermeabilization Solution with Saponin),可以用于免疫染色等多种原位检测时细胞样品、冰冻或石蜡切片的通透处理,有助于暴露抗原、核酸等作用靶点,使抗体、探针或标记物等更容易进入细胞内,从而确保染色等的检测效果。本产品为即用型工作液,可以直接使用,无需稀释。本产品通透能力较强,推荐用于各种常规的免疫荧光、免疫组化、免疫细胞化学、流式检测等中细胞的通透,特别适合细胞膜蛋白的检测。本产品不会溶解细胞膜,不会影响流式检测时的光散射。但本产品不推荐用于检测细胞核及线粒体内的蛋白。对于通透效果要求不高的情况,也可以使用阿拉丁的免疫染色洗涤液或免疫染色通透液(Triton X-100)。对于通透要求比较高的情况,可以使用免疫染色强力通透液。对于凝集素(lectin)的检测推荐使用免疫染色通透液(Triton X-100) 或免疫染色强力通透液。通透液通常可以通过使用有机溶剂例如甲醇、丙酮等,或者使用去垢剂例如Triton X-100、Saponin等来实现对于细胞膜的通透。甲醇或丙酮等有机溶剂,一方面可以溶解细胞膜和核膜从而充分暴露细胞浆和细胞核中的目的蛋白等,另一方面也可以使细胞内的蛋白变性起到固定的作用。甲醇或丙酮等有机试剂操作比较简单,一步处理可以同时实现固定和通透,缺点是膜蛋白也会被溶解掉并且有些蛋白变性后不利于后续的检测,因此有机溶剂使用相对较少,仅用于一些要求比较粗糙的检测。Triton X-100是常用的通透试剂,可以通透细胞膜和核膜,作用的原理也是非特异性地溶解细胞膜,因此其缺点是不利于膜蛋白的检测,但当使用多聚甲醛进行交联和固定后,相当一部分膜蛋白会被交联固定,从而不会被Triton X-100溶解,因此后续仍然能检测到。Saponin可以特异性地溶解细胞膜中的胆固醇,从而实现在细胞膜上选择性地打孔,其优点是适合于细胞膜蛋白的检测,特别适合通过流式细胞仪检测细胞膜上的标志性蛋白,缺点是对于一些胆固醇含量低的细胞通透效果差,通透效果弱于Triton X-100和有机溶剂,并且不能通透胆固醇含量很低的核膜和线粒体膜。对于凝集素(lectin)的检测,含Triton X-100等非特异性去垢剂的通透液的效果显著优于主要含Saponin的通透液。本免疫染色通透液(Saponin)中含有Saponin等去垢剂,配制在PBS中。经本免疫染色通透液(Saponin)处理的样品,对于细胞膜目的蛋白的免疫染色检测结果表明,染色效果与常规的通透液相比基本一致或有显著增强。按照每个样品需要0.1或1毫升免疫染色通透液(Saponin)计算,一个100ml包装的本产品可以通透1000个或100个样品,一个500ml包装的本产品可以通透5000个或500个样品。


注意事项

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.对于切片,在固定、洗涤结束后,每样滴50-100μl免疫染色通透液(Saponin),也可以在染色缸中完全浸没进行通透。对于细胞样品,在固定、洗涤结束后,按照六孔板每孔加入1毫升免疫染色通透液(Saponin),其它多孔板参考该比例加入。对于其它样品,加入免疫染色通透液(Saponin)的量以充分盖住样品为准。2.通常在免疫染色通透液(Saponin)中室温孵育5-10分钟,即可完成通透。对于较难通透的样品或者要求通透特别充分的情况,可以在免疫染色通透液(Saponin)中室温孵育10-30分钟。

Immunostaining Permeabilization Buffer with Saponin can be used for permeabilization of cell samples, frozen or paraffin sections in in situ assays such as immunostaining to expose the detection targets such as antigens and nucleic acids, allowing for efficient target labeling by antibodies, probes, or other markers. This ready-to-use product can be used directly without any extra preparations.This product has strong permeability and is recommended for cell permeabilization in various routine immunofluorescence, immunohistochemistry, immunocytochemistry, and flow cytometry assays, etc. It is especially suitable for detection of cell membrane proteins. This product does not dissolve cell membrane nor affect the light scattering during flow cytometry assay. However, this product is not recommended for detection of nuclear and mitochondrial proteins. For low permeation requirements, Aladdin's Immunostaining Wash Buffer or Immunostaining Permeabilization Buffer with Triton X-100 can also be used. For higher permeation requirements, Enhanced Immunostaining Permeabilization Buffer can be used. For lectin detection, Immunostaining Permeabilization Buffer with Triton X-100 or Enhanced Immunostaining Permeabilization Buffer is recommended.Permeabilization of cell membranes is usually achieved by using organic solvents such as methanol and acetone, or by using detergents such as Triton X-100 and Saponin. Organic solvents can lyse cell membrane and nuclear membrane to fully expose target proteins in cell plasma and nucleus, and also denature proteins in the meantime as a fixative. The disadvantage is that membrane proteins are also dissolved and some proteins are denatured, which is unfavorable sometimes for downstream applications. Therefore, organic solvents are rarely used except for some rough assays. Triton X-100 is a commonly used permeabilizing reagent that can permeate cell and nuclear membranes by non-specific lysis of cell membranes, thus its disadvantage is that it is not conducive for detection of membrane proteins. However, after cross-linking and fixation with paraformaldehyde, a significant portion of membrane proteins will not be lysed by Triton X-100 and can be detected subsequently. Saponin specifically solubilizes cholesterol in cell membranes and thus enables selective perforation on cell membranes. Its advantage is that it is suitable for examining cell membrane proteins, especially signature proteins on cell membranes by flow cytometry. Its disadvantage is that it is poorly permeable to cells with low cholesterol content, with weaker permeability than Triton X-100 and organic solvents, and cannot penetrate nuclear and mitochondrial membranes that have very low cholesterol content. For the detection of lectin, permeabilization solutions containing non-specific detergents such as Triton X-100 are significantly more effective than those containing Saponin mainly.This product contains Saponin and other detergents and is prepared in PBS. Immunostaining assay results of target cell membrane proteins in samples treated with this product is basically the same or even superior to that obtained with conventional permeabilization solutions.


Precautions

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. For slice sections, after fixation and washing, add 50-100μl of this product per sample, or permeabilize by complete submersion in this product in a staining jar. For cell samples, after fixation and washing, add 1ml of this product per well for six-well plates, or refer to this ratio for cells in other multi-well plates. For other samples, add sufficient Immunostaining Permeabilization Buffer to cover samples completely.2. Permeabilization can usually be completed by incubation at room temperature for 5-10 minutes. For samples that are more difficult to permeate or require full permeation particularly, incubate for 10-30 minutes at room temperature.


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