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Miracle-Star™ Western Blot Detection System

Cat.No Capacity Inquire
16028 200 ㎖ Inquire
PRODUCT INFORMATION
Description

High intensity, fast reaction, high sensitivity Chemiluminescence Western Blot products

  • • Able to detect pico-grams(pg) level of antigen on membrane blot.
  • • High Signal Intensity – Generate high contrast signals
  • • Duration Time - Maintain stable luminescence for 2 hours after reaction
  • • High reactivity - Excellent chemiluminescence reaction can be obtained by incubation for 1 to 5 min
  • • Economical - Highly reactive (with HRP) products can reduce the use of secondary antibodies

Miracle-Star™ Western Blot Detection System is able to detect specific antigen(protein) by chemiluminescence method using HRP(Horse radish peroxidase) with powerful signal intensity and high sensitivity. Western Blot Detection is common and essential method for protein expression research. The procedure is as follows. First, implement electrophoresis proteins including specific antigen by SDS-PAGE and then transfer to membrane like PVD and NC. After that, block the proteins that are not binding with membrane by BSA, skim milk or Gelatin solution and specific antigen(protein) binds with specific antibody. The Fc region of antibody binds with secondary antibody. At the secondary antibody, AP (Alkaline phosphatase) or HRP (Horse radish peroxidase) is binding.

Miracle-Star™ Western Blot Detection System reacts with secondary antibody which is binding with HRP and make chemiluminescence reaction. HRP binding with secondary antibody resolves Hydrogen peroxide included in Enhancer Solution of Miracle-Star™ Western Blot Detection System and Luminol is oxidized by oxygen isolated from Hydrogen peroxide and make strong blue-chemiluminescence. In the darkroom, exposure the chemiluminescence reaction to the X-ray Film and develop and print out. The band is detected where the specific antigen binds with antibody. It is very useful method for researching specific protein expression. Miracle-Star™TM Western Blot Detection System is composed Substrate Solution which included Luminol caused of chemiluminescence and Enhancer Solution which promotes strong luminescence. For using, mix each solution at a 1:1 ratio.

Protein detection process of Miracle-Star™ Western Blot Detection System

Miracle-StarTM Western Blot Detection System is upgraded initial intensity an duration time and it is optimized to detect traces of proteins effectively. Also, it is maintained for 1 year as the best quality by controlling reduction of Hydrogen peroxide and improving stability.

Applications
  • 01Western Blotting using chemiluminescence
  • 02Any Blotting methods (Southern, Northern) which use biotin-avidin-HRP
Kit Contents
Contents 16028
Substrate Solution 100ml × 1 ea
Enhancer Solution 100ml × 1 ea
Manual 1 ea
Technical Data

High Sensitivity

Human actin protein was isolated by western blot and compare the sensitivity to old version product. As the result, the intensity of the signal was improved and even after 1 hour from the reaction, we could see a higher signal than the previous product.

* Secondary antibody: 1/10,000 diluted, Reaction time : 1min, Exposed time : 1min
lane 1; 250 pg of antigen, lane 2; 125 pg of antigen, lane 3; 62.5 pg of antigen, lane 4; 31.3 pg of antigen, lane 5; 15.6 pg of antigen, lane 6; 7.8 pg of antigen, lane 7; 3.9 pg of antigen, lane 8; 2.0 pg of antigen.

Fast Reaction

It was confirmed that the chemiluminescence reaction by luminol was optimized and showed high reactivity within a short time. Due to its improved reactivity, it is possible to detect clear target proteins even with low amounts of antibodies, thus making economic Western Blot possible.

* Reaction time : 1min, and expose after 1 hour Exposed time : 1min
Lane M; Western Protein Marker, lane 1; 250 pg of antigen, lane 2; 125 pg of antigen, lane 3; 62.5 pg of antigen, lane 4; 31.3 pg of antigen, lane 5; 15.6 pg of antigen, lane 6; 7.8 pg of antigen, lane 7; 3.9 pg of antigen, lane 8; 2.0 pg of antigen.

TroubleShooting Guide
QThe initial sensitivity is too strong to create a white spot on the band. Is there a way to reduce this part?
AThe strong sensitivity may be due to the high amount of reactive protein or the high sensitivity of the antibody. Therefore, first try reducing the amount of initial loading sample. In addition, dilute the primary or secondary antibody, reduce the exposure time, increase the washing time after antibody binding, and reduce the intensity of the reaction if unnecessary antibody is removed.
QWhy does the back ground appear?
AIn the case of the back ground, if there is a lot of residue left after the solution treatment on the membrane, it may occur with the reaction with extra antibody. Reducing the amount of secondary antibody and increasing the washing time will give you good results. Also, please reduce the exposure time to 1 minute or 30 seconds.
QIs it possible to use the same method to detect using LAS4000 machine?
AOur products are applicable to machines and can be used in exactly the same way as X-ray films. If you use the same chemiluminescence detection method as the X-ray photoluminescence method in LAS equipment, you will have no problem confirming the results.
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