admin 管理员组文章数量: 1184232
2024年12月29日发(作者：web前端开发个人博客)
7th Edition, revised in April, 2017
（FOR RESEARCH USE ONLY. DO NOT USE IT IN CLINICAL DIAGNOSIS !）
Excellent Chemiluminescent Substrate Kit
Catalog No:
E-IR-R301
50 mL/ 100 mL /250 mL/ 500 mL/ 1000 mL
This manual must be read attentively and completely before using this product.
If you have any problems, please contact our Technical Service Center for help.
Phone: 240-252-7368(USA) 240-252-7376(USA)
Email: ***************************
Website:
Please kindly provide us the lot number (on the outside of the box) of the kit for more efficient service.
Copyright ©2017-2018 Elabscience Biotechnology Inc. All Rights Reserved
7th Edition, revised in April, 2017
Description
Excellent Chemiluminescent Substrate is an enhanced chemiluminescent (ECL) substrate for horseradish
peroxidase (HRP) enzyme that provides low picogram detection of proteins in Western blot and ELISA
applications.
Excellent Chemiluminescent Substrate emits light for longer and chemiluminescent signal at more than
twice the intensity of other entry-level luminol-based horseradish peroxidase detection systems while
also maintaining sensitivity and low background. In addition, the range of the calibration curve is much
wider than other competitors. Also, the greater signal intensity is achieved using less primary and
secondary antibody than typical ECL substrates. All of this for about 30% less cost than comparable
products.
Product features
>
>
>
>
>
>
>
>
ECL substrate – An enhanced chemiluminescent substrate for detection of horseradish
peroxidase (HRP) activity of antibodies and other ELISA or Western blot probes.
Economy – Costs less per milliliter than other similarly sensitive chemiluminescent substrates.
Picogram sensitivity – Highly sensitive for the rapid development of a wide range of protein
levels.
Stable light output – Long-duration signal makes it possible to make multiple exposures to film.
High intensity – Signal is twice as strong as output from other luminol-based systems.
Excellent stability – 8-hour working solution stability; 1-year kit stability at 2~8℃.
Saves antibody – Requires much less (more dilute) primary and secondary antibodies per blot.
Wide range of the calibration curve –Calibration curves range from 4.00–8000 pg, R2 is 0.992.
Kit components
Item
Substrate A
Substrate B
Product Description
50 mL
25mL
25 mL
100 mL
50 mL
50 mL
250 mL
125 mL
125 mL
1 copy
500 mL
250 mL
250 mL
1000 mL
500 mL
500 mL
2~8℃
Storage
2
7th Edition, revised in April, 2017
Detailed Western Blotting Procedure
1. Remove the blot from the transfer apparatus and block nonspecific sites with Blocking Reagent
for 20-60 minutes at room temperature (RT) with shaking. For best results, block for 1 hour at RT.
Note: Avoid using milk as a blocking reagent when using Avidin/Biotin systems because
milk contains variable amounts of endogenous biotin.
2. Remove the Blocking Reagent and add the appropriate primary antibody dilution. Incubate the blot
for 1 hour with shaking. If desired, blots may be incubated with primary antibody overnight at 2-
8°C.
3. Wash the membrane by suspending it in Wash Buffer and agitating for ≥ 5 minutes. Replace Wash
Buffer at least 4-6 times. Increasing the wash buffer volume and/or the number of washes may help
reducing background.
Note: Briefly rinsing membrane in wash buffer before incubation will increase wash
efficiency.
4. Incubate the blot with the appropriate HRP-conjugate dilution for 1 hour at RT with shaking.
5. Repeat Step 3 to remove non-bound HRP-conjugate.
Note: Membrane must be thoroughly washed after incubation with the HRP-conjugate.
6. Prepare Working Solution by mixing equal parts of the Substrate A and Substrate B. Use 0.1 mL
Working Solution per cm
2
of membrane. The Working Solution is stable for 8 hours at RT.
Note: Exposure to the sun or any other intense light can harm the Working Solution. For
best results keep the Working Solution in an amber bottle and avoid prolonged exposure to
any intense light. Typical laboratory lighting will not harm the Working Solution.
7. Incubate the blot with Working Solution for 5 minutes.
8. Remove the blot from Working Solution and place it in a plastic membrane protector, a plastic
sheet protector or plastic wrap may be used. Use an absorbent apparatus to remove excess liquid
and to carefully press out any bubbles between the blot and surface of the membrane protector.
9. Place the protected membrane in a film cassette with the protein side facing up. Turn off all lights
except those appropriate for film exposure (e.g., a red safelight).
Note: Film must remain dry during exposure. For optimal results, perform the following
precautions:
• Make sure excess substrate is removed from the membrane and the membrane protector.
• Use gloves during the entire film-handling process.
• Never place a blot on developed film, as there may be chemicals on the film that will reduce
signal.
3
7th Edition, revised in April, 2017
10. Carefully place a piece of film on top of the membrane. A recommended first exposure time is 60
seconds. Exposure time may be varied to achieve optimal results. Enhanced or pre-flashed film is
unnecessary.
Note: The exposure time may be varied to achieve optimal results. If the signal is too intense,
reduce exposure time or optimize the system by decreasing the antigen and/or antibody
concentrations.
Light emission is most intense during the first 5-30 minutes after substrate incubation. Light
emission will continue for several hours, but will decrease with time. Longer exposure times
may be necessary as the blot ages.
Attention: Light emission is intense and any movement between the film and membrane may
cause artifacts on the film.
Copyright ©2017-2018 Elabscience Biotechnology Inc. All Rights Reserved
4
版权声明：本文标题：Elabscience Excellent Chemiluminescent Substrate K 内容由网友自发贡献，该文观点仅代表作者本人， 转载请联系作者并注明出处：http://www.roclinux.cn/p/1735521090a1673850.html， 本站仅提供信息存储空间服务，不拥有所有权，不承担相关法律责任。如发现本站有涉嫌抄袭侵权/违法违规的内容，一经查实，本站将立刻删除。
发表评论