产品
- 产品介绍
- 相关文献
- 实验结果
- 实验条件
- 常见问题
- Catalog No.
ES-0006
- Size
100mL/500mL
- Form
liquid
- Compatibility
Nitrocellulose membrane and PVDF membrane
- Detection method
Chemiluminescence
- Substrate type
HRP (horseradish peroxidase) substrate
- Storage
It is recommended to store the product at 2–8 °C. Keep the container tightly closed and keep away from heat and light. The product is stable for a minimum of 18 months when stored in the dark at 2–8 °C.
- Background
Enhanced chemiluminescence (ECL) substrate is a two-components product for the detection of horseradish peroxidase. This substrate is a highly sensitive, luminol-based solution that can be used for any blotting applications that utilize horseradish peroxidase (HRP)-conjugates, with minimal background interference.
- Components
Item
100mL
500mL
Solution A
50mL
250mL
Solution B
50mL
250mL
- Procedure
In order to prevent contamination, do not pour used solution back into the substrate bottle. Use clean gloves and forceps; do not use rusted forceps. Analytes can be applied to membranes as a dot blot or by gel transfer. Blotting conditions should be optimized for each assay system.
1.Preparation of working solution: Mix an appropriate amount of solution A and solution B in equal volume, and store at room temperature for later use.Working solution should be prepared before testing.
2.After incubation with Western secondary antibody and washing for several times, remove the membrane with WB special forceps, slightly absorb excess liquid with absorbent paper (do not touch the protein surface of the membrane), and then place it on a clean plastic wrap.
3.According to the size of the membrane, add 1 mL of working solution to the membrane per 10 square centimeters, make sure that the working solution is evenly covered on the membrane, and leave for 1-2 minutes..
4.Discard the working solution, and slightly absorb the excess liquid with a absorbent paper.The membrane is placed between two pieces of plastic wrap and then tested by a tablet compression or chemiluminescence imager.
5.Tablet compression test: fix the film in the tablet clip.Pressing the tablet in the darkroom for 1 minute, immediately develop and fix the film, and adjust the pressing time according to the results.Or directly press the tablet for 30 seconds, 1, 3, 5 minutes, and then develop and fix the observation results together.
6.Chemiluminescence imager test: place the film in the chemiluminescence imager, refer to the instrument manual for testing.
- Precautions and Disclaimer
1.The spearhead must be replaced in the process of absorbing liquid A and liquid B. The mutual contamination of liquid A and liquid B will lead to gradual failure of liquid A or liquid B, affecting the subsequent use effect.
2.After using the solution, please close the bottle cap to prevent failure.Liquid B, in particular, contains oxidant, which is easy to be reduced and ineffective.
3.Both solution A and solution B are harmful to human body. Please be careful when handling and pay attention to effective protection to avoid direct contact with human body or inhalation.
4.This product is for R&D use only, not for drug, household, or other uses.
- 期刊名：
- Redox Biology
- 作者：
- 山东第一医科大学
- 发布日期：
- 2024-01-26
- 引用产品：
- 期刊名：
- Biochemical Pharmacology
- 作者：
- 复旦大学附属闵行医院
- 发布日期：
- 2025-04-18
- 引用产品：
- 期刊名：
- Current Molecular Medicine
- 作者：
- 河北北方学院附属第一医院
- 发布日期：
- 2024-01-26
- 引用产品：
- 期刊名：
- Applied Entomology and Zoology
- 作者：
- 遵义医科大学生物化学与分子生物学
- 发布日期：
- 2023-07-28
- 引用产品：
抗体可以回收利用几次？
一般抗体不推荐回收利用，抗体使用之后缓冲体系已经发生改变，不同客户在回收抗体的保存条件上也会有差异，所以抗体回收使用效果无法保证。
FineTest公司也做过一批抗体回收验证测试，测试结果显示不同抗体可回收次数不同，一般效价越高的抗体，可重复使用的次数越多，客户可根据实验情况来确定。
注：我们将孵育完毕后剩余的抗体回收到离心管中置于4℃保存，效价高的抗体可至少保存1周，重复利用3次左右。
FineTest公司抗体buffer成分是什么？
FineTest抗体成分一般是PBS含proclin300或sodium azide、BSA、50%甘油；也有一些是PBS含proclin300或sodium azide，50%甘油。防腐剂proclin300或sodium azide是常用的防腐剂，广泛应用于实验室和工业中。
FineTest公司抗体的保存温度和时间？
大部分抗体是放在-20℃保存，直标流式抗体需要放在2-8℃保存，质保期一年，质保期内抗体出现售后，可进行退货或换货。
一般抗体超过一年质保期仍旧可以使用，我们可以提供技术支持服务。
FineTest公司抗体使用需要稀释吗？用什么溶液稀释？
除了直标流式抗体为即用型，不用稀释。其他抗体一般为浓缩型，使用时需要按照说明书建议稀释比例稀释。
不同实验使用的稀释液不同，用PBST, TBST, 抗体封闭液等常用抗体稀释液均可。
免疫组化实验抗体的修复方式？
常用修复液：Tris-EDTA缓冲液（Tris-EDTA Buffer, pH 9.0）；柠檬酸盐缓冲液（Citrate Buffer, pH 6.0）
热修复：
方法1：沸水浴修复，将盛有修复液和玻片的烧杯置于沸水浴环境，保持外部沸腾状态15min，自然冷却至室温；
方法2：微波修复，将盛有修复液和玻片的烧杯置于微波炉中，高火5min，停火3min，中火5min，自然冷却至室温。
如何选择二抗？
（1）二抗是和一抗反应的，所以二抗必须是抗一抗宿主物种，比如一抗来源于兔，对应二抗则选择抗兔二抗，比如羊抗兔或驴抗兔。
（2）根据实验类型选择二抗偶联物，比如做ELISA, 蛋白质印迹，IHC实验，常用酶联二抗，如HRP，AP等标记。而免疫荧光实验，流式细胞术实验中，则用荧光蛋白或染料标记，比如FITC，Cy3等。
