2013年3月18日 星期一

TOYOBO Can Get Signal 減少抗體使用 增加wesrern 訊號


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Can Get Signal   三大優點
Can Get Signal  增強免疫分析迅號的數十倍，保持較低的背景迅號
Can Get Signal  可以用在已標記過氧化物酶HRP或鹼性磷酸酶AP之相關抗體等
Can Get Signal  可以直接使用，無需稀釋
 
 


Can Get Signal®
Immunoreaction Enhancer Solution

Code No. NKB-101T 50 ml x 2
NKB-101 250 ml x 2
NKB-201 250 ml x 1
NKB-301 250 ml x 1
 

Description

Can Get Signal® is a solution containing an accelerator for antigen-antibody reactions. This reagent improves sensitivity, specificity, and signal-to-noise ration (S/N) for Western blotting, dot blotting, enzyme-linked immunosorbent assay (ELISA), etc. Solutions 1 and 2 refer to the reactions of the primary and secondary antibodies, respectively.


Features

  • Enhances immunoassay signals up to several dozen times by maintaining low background signals.
  • Can be used in combination with secondary antibodies labeled with peroxidase or alkaline phosphatase, etc.
  • Can be used directly without dilution (ready-to-use).


Applications

  • Western blotting, dot blotting
  • Enzyme-linked immunosorbent assay (ELISA),


Storage condition

Store at 4ºC


Components

This kit includes the following components. All reagents should be stored at 4ºC, and protected from light.

Reagent NameCode No.
NKB-101TNKB-101NKB-201NKB-301
Solution 1
for primary antibody		50 ml250 ml250 ml-
Solution 2
for secondary antibody		50 ml250 ml-250 ml


Typical reaction flow

Flow chart of western blotting with Can Get Signal®
Flow chart of ELISA with Can Get Signal®


Application data

Experiment 1. Detection of phosphorylated proteins by Western blotting
Phosphorylated Akt and ERK were detected by Western blotting analysis using Can Get Signal® and a conventional method (TBS-T). As a result, the signal intensities of the target bands obtained with Can Get Signal® were greater than those of the conventional method. The background level of the experiment with Can Get Signal®was also significantly lower than that of the conventional method. The results suggest that Can Get Signal® improves the sensitivity and specificity of Western blotting analysis.

Sample:Cultured bovine adrenal medulla cells

1. Control (H2O)
2. Insulin (1 nM, stimulated for 5 min)
3. Insulin (10 nM, stimulated for 5 min)
4. Insulin (100 nM, stimulated for 5 min)

Antibodies:
<p-Akt>
Primary antibody:Anti Phospho-Akt rabbit polyclonal antibody (1:2,000 dilutioin)
Secondary antibody:Anti rabbit-HRP antibody (1:20,000 dilution)

<p-ERK>
Primary antibody:Anti Phospho-ERK monoclonal antibody(1:2,000 dilutioin)
Secondary antibody:Anti mouse-HRP antibody (1:20,000 dilution)

Fig.1 Detection of phosphorylated protein kinases (p-Akt, p-ERK1 and p-ERK2) by Western blotting with Can Get Signal® and a conventional method
*The data was kindly provided by Dr. Yanagita from the Department of Pharmacology, Faculty of Medicine, University of Miyazaki.
Experiment 2. Detection of His-tagged proteins by Western blotting
His-tagged recombinant proteins were detected with Can Get Signal® and a conventional method (TBS-T). Can Get Signal® showed excellent greater sensitivity than the conventional method.

Antibodies:
Primary antibody:Anti his-tag rabbit polyclonal antibody (1:2,000 dilutioin)
Secondary antibody:Anti rabbit-IgG-HRP antibody (1:20,000 dilution)
Fig.2 Detection of His-tagged proteins by Western blotting
Experiment 3. Detection of His-tagged proteins by ELISA
Sandwich ELISA (solid phase antibody: anti-ERK2 monoclonal antibody, primary antibody: anti-His tag polyclonal antibody, secondary antibody: anti-rabbit IgG-HRP antibody) was performed to detect his-tagged human MAP kinase (His-ERK2) synthesized by a cell-free protein synthesis system. Can Get Signal® showed an excellent quantitative curve as a function of antigen concentration whereas the conventional method with TBS-T resulted in low signals.

Fig.3 Detection of His-tagged proteins by sandwich ELISA





萊恩生物科技有限公司 http://biolion.blogspot.tw
台中專員 陳俊豪  0937-229803   04-25239639

 

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