
- Overview
- Data/Specifications
- Literature/Support
- How To Use
- Related Products
Overview
Isotype controls are most commonly used in flow cytometry and immunohistochemistry experiments as negative controls that measure the level of non-specific background signal caused by primary antibodies. Background signal is due to Igs binding to non-specifcally to Fc receptors present on a cell surface (ex. mice antibodies binding to human leukocytes non-specifically in which investigators would want to use a mouse isotype control when working with human cells or tissues).
Data/Specifications
Ig Isotype: Rat IgG1k
Specifcity:Keyhole limpet hemocyanin (KLH)
Form:This product supplied as 0.5 mg of purified immunoglobulin in 1.0 mL of borate buffered saline, pH 8.2.
Storage:Store at 2 – 8 °C
Literature/Support
PDF Insert
How To Use
Applications:
- Used for Flow cytometry.
- Used for Enzyme-Linked–Immunosorbent-Assay (ELISA): recommended dilution of 1 ug/mL
- Used for Flourescent-Linked-Immunosorbent-Assay (FLISA).
Note: Since applications vary, each investigator should determine the optimum working dilutions of the product that is appropriate for their specific needs
Related Products
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我要做的两个目的蛋白,分子量一个是140的,另一个是36,内参用的GAPDH。这几天刚刚要做小鼠组织的蛋白,就把新提取的组织蛋白和之前做过的细胞蛋白一起上了,细胞和组织的各上了3个孔。
电泳转膜孵育等条件都是以前做成熟的
之后显影,细胞蛋白的,两个目的和内参都出得挺好;但是小鼠的组织蛋白只有分子量140的那个显出条带了,36的那个显出来的像一片水渍一样,斑片状的黑,内参有时也这样、有时干脆什么都不显。不知道这是什么原因...按说细胞同样的蛋白做出来,WB的各个步骤应该不会有太大问题的。
有点怀疑蛋白提取的不好,我提组织蛋白的的操作是在冰上进行的,裂解液也加有PMSF,用BCA法测得的蛋白浓度大概也有3~5mg/ml。
在这里求助各位高手~望给我指点一下~不胜感激啊!
去垢剂作为添加剂生长蛋白质晶体是一项很有用的技术。非离子和两性离子结晶剂在膜蛋白结晶中已经得到很好应用,目前已经变成常规的实验手段。去垢剂在一些可溶蛋白中也起到了一定的作用,提高了晶体的质量和结果的可重复性 ,促进了单晶的生长, 但具体的影响机制尚不明确。其可能原因是去垢剂的引入使结晶液滴中的蛋白分子更具有亲水性,使溶液中同质蛋白质分子增加。向左转|向右转
如要膜蛋白保持原来的结构应采用:非离子型去垢剂.
表面活性剂:通常由一个极性头和一个非极性尾组成,用于去除难溶于水的污渍
漂白剂:通常含次氯酸盐成分,原理为其氧化性
稀盐酸:用于去除碱性的污渍,如马桶中的结块
碳酸钠溶液:用于去除油渍,其原理为酯在碱溶液中水解,参见皂化反应

