本产品为辣根过氧化物酶标记山羊抗小鼠IgG(IgG-HRP),由亲和纯化的高纯度羊抗小鼠IgG和高纯度的辣根过氧化物酶交联而成,用于Western、ELISA和IHC等的二抗。HRP,即horseradish peroxidase,中文名为辣根过氧化物酶,可以在Western检测时催化ECL类试剂(如ECL、BeyoECL等)产生化学发光,可以在ELISA时催化TMB产生蓝色,也可以在IHC或Western blot检测时催化DAB产生棕色沉淀。
本辣根过氧化物酶标记山羊抗小鼠IgG用于WB、ELISA和IHC的推荐稀释比例参考下表,实际实验操作过程中需根据抗原和抗体的具体情况适当调节二抗的稀释比例。
应用范围:
抗原决定族的抗原性分析鉴定,各种抗原、抗体的定量、定性及定位分析测定等。
工作中易产生的问题:
1. 包被抗体或抗原不适,易产生阳性值较小,或产生变异,即跳管现象。
2. 用抗血清不纯化,或纯化程度太低的抗体包被,则阳性值较小,或没有阳性梯度。
3. 封闭不好或标记物过浓可产生对照较高。
保存条件:
-20℃保存,一年有效;4℃保存,1个月有效;避免反复冻融。
储存液:0.01M PBS(pH7.4) with BSA, Glycerol。
应用推荐稀释比如下:
来源 |
用途 |
WB |
ELISA |
IHC |
Goat |
WB, ELISA, IHC |
1:500`5000 |
1:1000`5000 |
1:100`1000 |
This product is horseradish peroxidase labeled goat anti mouse IgG (IgG-HRP), cross-linked from affinity purified high-purity goat anti mouse IgG and high-purity horseradish peroxidase, used for secondary antibodies such as Western blotting, ELISA, and IHC. HRP, Horseradish peroxidase, also known as horseradish peroxidase in Chinese, can catalyze ECL reagents (such as ECL, BeyoECL, etc.) to produce chemiluminescence in Western detection, catalyze TMB to produce blue in ELISA, and catalyze DAB to produce brown precipitate in IHC or Western blot detection.
The recommended dilution ratios for goat anti mouse IgG labeled with horseradish peroxidase for WB, ELISA, and IHC are shown in the table below. During the actual experimental operation, the dilution ratio of the secondary antibody should be adjusted appropriately according to the specific situation of the antigen and antibody.
Application scope:
Antigenicity analysis and identification of antigenic determinants, quantitative, qualitative, and positional analysis and determination of various antigens and antibodies.
Problems that are prone to occur in work:
1. Discomfort with antibodies or antigens can lead to small positive values or mutations, known as tube skipping.
If the antibody is not purified with anti serum or the purification level is too low, the positive value will be small or there will be no positive gradient.
3. Poor sealing or excessive concentration of markers can result in a higher control level.
Storage conditions:
Stored at -20 ℃, valid for one year; Stored at 4 ℃, valid for 1 month; Avoid repeated freezing and thawing.
Storage solution: 0.01M PBS (pH 7.4) with BSA, Glycerol.
The recommended dilution ratio for the application is as follows:
来源 |
用途 |
WB |
ELISA |
IHC |
Goat |
WB, ELISA, IHC |
1:500`5000 |
1:1000`5000 |
1:100`1000 |