The beta-galactosidase assay is commonly used as a reporter, to monitor transfection efficiency, and for promoter/enhancer studies. We offer two ways to detect LacZ gene activity: a colorimetric beta-galactosidase staining kit and a chemiluminescent beta-galactosidase assay.
The beta-galactosidase assay is commonly used as a reporter, to monitor transfection efficiency, and for promoter/enhancer studies. We offer two ways to detect LacZ gene activity: a colorimetric beta-galactosidase staining kit and a chemiluminescent beta-galactosidase assay.
- Colorimetric assay: The Beta-Galactosidase Staining Kit is a complete system for colorimetric detection of beta-galactosidase expression in mammalian cells. In this kit, beta-galactosidase (the product of the LacZ gene) catalyzes the hydrolysis of X-gal to produce a blue color that is easily visualized. Simply fix, wash, and incubate your cells with the X-Gal Staining Mix. Then use a light microscope to look for the blue stain.
- Chemiluminescent assay: The Luminescent Beta-Galactosidase Reporter System 3 (which includes vectors; see below) and Detection Kit II (reagents only) provide quantitative measurement of beta-gal activity using a chemiluminescent substrate. The chemiluminescent signal is stable over several hours and can be detected by a scintillation counter, tube and plate luminometers, or by x-ray film exposure.
LacZ vectors
Beta-galactosidase is the product of the LacZ gene. We offer three beta-gal mammalian reporter vectors containing the LacZ gene. The vectors provide maximal flexibility in analyzing the effects of cis-regulatory elements on gene expression. They can also be used to normalize transfection efficiencies.
- pBeta-Gal-Basic (sold separately as pLacZ-Basic) lacks eukaryotic promoter and enhancer sequences and can serve as a negative control or to clone promoters for characterization.
- pBeta-Gal-Control (sold separately as pLacZ-Control) contains the SV40 promoter and enhancer sequences and can be used as a positive control or as a reference for comparing the activities of promoters and enhancers.
- pCMV-LacZ is a constitutive mammalian reporter vector which expresses extremely high levels of beta-galactosidase from the human cytomegalovirus immediate early promoter (PCMV IE) and can be used as a reference or control plasmid. Beta-galactosidase activity can be detected with the Luminescent Beta-galactosidase Detection Kit II.
Complete beta-gal reporter system
The Luminescent Beta-galactosidase Reporter System 3 includes the chemiluminescent beta-galactosidase assay reagents plus the pBeta-Gal-Basic and pBeta-Gal-Control vectors.
ebiomall.com
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一般可与大鼠、小鼠、人、兔均有交叉反应,即可作为上述任一来源蛋白的内参。
Actin分子量为42kD,为肌动蛋白
GAPDH分子量为36kD,为磷酸脱氢酶
虽然没有具体用过检测这个抗原的抗体,但是如果希望抗体特异性好点,能检测丰度低的蛋白,还是选Abcam吧,Santa有时候全凭人品了,内参抗体什么的还能凑活,其他的比较勉强。
当然跟物种有关。
内参抗体:
Beta-Actin mAb (1C7) 细胞总蛋白 42 kD Abbkine A01010 小鼠源单克隆
Beta-Actin mAb (1C7) , HRP 细胞总蛋白 42 kD Abbkine A01015 小鼠源单克隆,HRP偶联
GAPDH mAb (2B5) 细胞总蛋白 36 kD Abbkine A01020 小鼠源单克隆
GAPDH mAb (2B5) , HRP 细胞总蛋白 36 kD Abbkine A01025 小鼠源单克隆,HRP偶联
Beta-Tubuline mAb (3G6) 细胞总蛋白 55 kD Abbkine A01030 小鼠源单克隆
Plant Actin mAb (3T3) 植物细胞总蛋白 42 kD Abbkine A01050 小鼠源单克隆
PCNA mAb (1D7) 细胞核蛋白 28 kD Abbkine A01040 小鼠源单克隆
COX IV mAb (14Y2) 细胞线粒体总蛋白 16 kD Abbkine A01060 小鼠源单克隆
Histone H3 mAb (2D10) 细胞核蛋白 18 kD Abbkine A01070 小鼠源单克隆
标签抗体(Tag Antibody)可用于检测各种商品化表达载体上的标签序列(如:Myc、Flag、His、GST、HA等),籍以分析目的蛋白的表达含量及其功能;
标签抗体:
Anti-Biotin Antibodies
Anti-Dye Antibodies
Anti-FITC Antibodies
Anti-Fluorescent Protein Antibodies
Anti-HRP Antibodies
Beta Galatosidase Antibodies
FLAG Tag Antibodies
GST Tag Antibodies
HA Tag Antibodies
His Tag Antibodies
Myc Tag Antibodies
TAP Tag Antibodies
V5 Tag Antibodies
高保真生物技术有限公司~实验整体外包服务商~
我觉得封闭这一步时间长短多少并不能减少背景…背景过深多半还是抗体浓度过大…不管一抗还是二抗过大都会使背景加深…一抗的浓度不一定是条带清晰就是最佳了…联系进一步优化一下
这个主要还是抗体的问题,WB最主要的还是要选择一个特异性较好的抗体,抗体是整个实验的重中之重
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