计算溶液所需的质量、体积或浓度。
通用型ELISA溶液套装（7组分）
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
通用型ELISA溶液套装(Universal ELISA Buffer Set)，是一种包含了双夹心ELISA等ELISA实验过程中常用的包被液、封闭液、洗涤液、通用稀释液、显色液和终止液的溶液套装，可满足常规ELISA的溶液需求。用户仅须自备ELISA检测用抗体(如使用生物素体系，须自备生物素检测相关试剂)和酶标板及封板膜(如需进行绝对定量，还须自备标准品)，就可以进行ELISA检测。
ELISA (Enzyme-Linked Immunosorbent Assay)即酶联免疫吸附试验，是一种结合抗原、抗体特异性反应和酶对底物高效催化作用的高敏感性免疫学实验技术，广泛用于生物学和医学科学等的许多领域。常见的ELISA有双抗体夹心ELISA (Sandwich ELISA)、竞争ELISA (Competitive ELISA)、直接ELISA (Direct ELISA)、间接ELISA (Indirect ELISA)等。双抗体夹心
ELISA，简称双夹心ELISA，是最常见的ELISA检测方式之一，可用于检测样品中靶蛋白等目标分子的浓度。
使用说明：
ELISA的步骤参考ELISA试剂盒说明书。以下为本试剂盒中各个溶液的常规使用方法，具体可根据实际情况进行条件的优化。
1. 包被液。
无须稀释或配制，直接使用。使用包被液将捕获抗体稀释至合适浓度，100μl/孔加入到空白酶标板中，4ºC孵育过夜，或者37ºC孵育2小时。
2. 封闭液。
无须稀释或配制，直接使用。包被完成后，用洗涤液(1X，配制方法见步骤3)洗涤3次。加入200μl/孔封闭液，室温封闭120分钟，甩去孔内液体，拍干后直接进行一抗孵育等后续操作；或室温封闭2小时，甩去孔内液体，拍干后置于37ºC倒扣烘干90分钟，即可作为预包被板使用。预包被板封装在含有干燥剂的密闭防潮袋中，或在其它适宜的保存环境中，通常置于4ºC可保存约六个月。
3. 洗涤液(10X)。
a. 以配制1升洗涤液(1X)为例：将10ml洗涤液(10X)，倒入到一洁净的量筒中，加入蒸馏水混匀并定容到100mL后即为洗涤液(1X)，即可用于ELISA的洗涤。没有用完的洗涤液可以室温保存，通常1-2周内可正常使用，如需长时间保存，请将未使用完毕的洗涤液在4ºC保存，通常2个月内可正常使用。如果发现洗涤液出现浑浊、沉淀等异常现象，宜废弃。
b. 自动洗板机或手工洗板：每孔洗涤液为300μl，注入与吸出间隔15-30秒。洗板3-5次。最后一次洗板完成后将板倒扣在厚吸水纸上适当用力拍干。然后进行后续的操作。如果按照上述洗涤条件ELISA结果空白孔的信号值仍然较高，可以适当延长洗涤时间，并增加洗涤次数。通常，延长洗涤时间或增加洗涤次数不会对ELISA结果产生负面影响。
4.显色液。
将显色液A、B、C按照5:4:1比例配制。在SA-HRP或其它HRP标记抗体孵育后，用洗涤液(1X)洗涤3-5次，洗涤完毕后，去除洗涤液，加入100μl/孔显色液，室温显色5-30分钟，直至显色至预期深浅。本产品反应速度较快，如有必要可适当缩短反应时间。
6. 终止液。
无须稀释或配制，直接使用。加入100μl/孔终止液终止反应，随后在测定吸光度。使用终止液终止反应后在1小时内吸光度保持稳定。
注意事项：
1. 本套装产品仅提供各种溶液，高结合力酶标板、封板膜、封板膜刮板等需要自备)。
2. 使用前，将试剂盒内的溶液从4ºC冰箱取出，恢复至室温使用。
3. 使用时建议将各组分吸出或倒出所需用量使用，尽量避免直接从瓶中进行移液操作；未使用的多余溶液请勿倒回。
4. 洗涤液(10X)在低温下可能有结晶，如果发现有结晶，请室温水浴加热使结晶完全溶解后再配制工作液。 洗涤液(10X)配制成1X洗涤液后，宜4ºC保存，并在1-2个月内用完，如果室温保存，宜在1-2周内用完。
5. 如果发现TMB显色液出现混浊或颜色变成蓝色，应该停止使用。
6. 加入TMB显色液或者终止液后，如果有沉淀生成，说明样品中目的蛋白的含量过高，建议适当稀释后再进行测试。
7. 终止液有腐蚀性，操作时请小心，并确保有效防护以避免直接接触人体，并须注意避免腐蚀其它物品。
8. 本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
9. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Universal ELISA Buffer Set is a comprehensive solution kit containing commonly used buffers and reagents for ELISA procedures, including sandwich ELISA. The set includes coating buffer, blocking buffer, wash buffer, universal dilution buffer, chromogenic substrate, and stop solution, meeting the standard solution requirements for routine ELISA. Users only need to provide the ELISA detection antibodies (and biotin detection reagents if using a biotin system), microplates, and sealing films (and standards if absolute quantification is required) to perform ELISA.
ELISA (Enzyme-Linked Immunosorbent Assay) is a highly sensitive immunological technique that combines the specific reaction between antigen and antibody with the efficient catalytic action of an enzyme on its substrate. It is widely used across many fields in biology and medical science. Common types of ELISA include the double antibody sandwich ELISA (Sandwich ELISA), competitive ELISA, direct ELISA, and indirect ELISA. The double antibody sandwich ELISA, often abbreviated as sandwich ELISA, is one of the most common ELISA formats and can be used to determine the concentration of target molecules, such as specific proteins, in a sample.
Instructions for Use
Refer to the specific ELISA kit instructions for detailed procedural steps. The following describes the general for each solution in this set; conditions can be optimized according to actual experimental needs.
1. Coating Buffer
Use directly without dilution or reconstitution.Dilute the capture antibody to an appropriate concentration using the Coating Buffer.Add 100 µL per well to a blank microplate.Incubate overnight at 4°C, or for 2 hours at 37°C.
2. Blocking Buffer
Use directly without dilution or reconstitution.After coating, wash the plate 3 times with Wash Buffer (1X, preparation see step 3).Add 200 µL of Blocking Buffer per well and incubate at room temperature for 120 minutes.Discard the liquid, blot the plate dry, and proceed directly to subsequent steps such as primary antibody incubation.For preparing pre-coated plates: After room temperature blocking for 2 hours, discard the liquid and blot dry. Invert and dry the plate at 37°C for 90 minutes. Pre-coated plates can be sealed in a moisture-proof bag with desiccant or stored in other suitable conditions. Typically, they can be stored at 4°C for approximately six months.
3. Wash Buffer (10X)
a. Preparation of Working Solution (1X).To prepare 1 liter of Wash Buffer (1X) as an example: Pipette 10 mL of Wash Buffer (10X) into a clean measuring cylinder. Add distilled water, mix, and bring to a final volume of 100 mL. This is now ready-to-use Wash Buffer (1X).Unused prepared Wash Buffer (1X) can be stored at room temperature and is typically stable for 1-2 weeks. For longer storage, keep unused Wash Buffer (1X) at 4°C, where it is typically stable for up to 2 months.Discard the buffer if turbidity or precipitation is observed.
b. Automatic plate washer or manual plate washing: Add 300 μl of washing buffer to each well and allow an interval of 15–30 seconds between dispensing and aspiration. Wash the plate 3–5 times. After the final wash, invert the plate and pat it firmly on thick absorbent paper to dry completely. Proceed with subsequent steps.If the signal value of blank wells in the ELISA remains high under the above washing conditions, appropriately extend the washing time and increase the number of washing cycles. Generally, extending the washing time or increasing the washing frequency will not exert a negative impact on the ELISA results.
4. Color Development Solution
Prepare the working solution by mixing Components A, B, and C at a 5:4:1 volume ratio.
After incubation with SA-HRP or other HRP-conjugated antibodies, wash the plate 3-5 times with Wash Buffer (1X).After the final wash and removal of buffer, add 100 µL of the prepared Color Development Solution per well.ncubate at room temperature for 5-30 minutes, or until the desired color intensity is achieved.Note: This product reacts rapidly. The incubation time can be shortened if necessary.
5. Stop Solution
Use directly without dilution or reconstitution.Add 100 µL of Stop Solution per well to terminate the reaction.Read the absorbance promptly thereafter. The absorbance typically remains stable for up to 1 hour after stopping the reaction.
Notes:
1. This set provides buffers and solutions only. High-binding microplates, plate sealers, sealer scrapers, etc., must be supplied by the user.
2. Before use, allow all solutions to equilibrate to room temperature after removing them from 4°C storage.
3. It is recommended to aliquot the required volume for use. Avoid pipetting directly from the stock bottles. Do not return unused solution to the original container.
4. Wash Buffer (10X) may crystallize at low temperatures. If crystals are observed, warm the bottle in a room temperature water bath until crystals are completely dissolved before preparing the working solution.Prepared Wash Buffer (1X) is best stored at 4°C and used within 1-2 months. If stored at room temperature, use within 1-2 weeks.
5. If the TMB Color Development Solution appears turbid or turns blue, it should not be used.
6. If precipitation forms after adding the TMB Color Development Solution or Stop Solution, it may indicate an excessively high concentration of the target protein in the sample. It is recommended to dilute the sample and repeat the test.
7. Caution: Stop Solution is corrosive. Handle with care. Wear appropriate personal protective equipment to avoid contact with skin or eyes. Take care to avoid corrosion of other materials.
8. This product is for research use only by qualified professionals. It is not intended for diagnostic or therapeutic use, nor for use in food or drugs. Do not store in ordinary household areas.
9. For your safety and health, wear a lab coat and disposable gloves during operation.
规格
别名
ELISA辅助组分试剂盒 | ELISA补充溶液套装
英文别名
ELISA Ancillary Reagent Kit | ELISA Supplemental Solution Set | ELISA universal kit
规格或纯度
BioReagent, 用于蛋白分析, 无菌, for ELISA
英文名称
Universal ELISA Buffer Set (7 Components)
应用
ELISA
检测物种
抗原; 抗体
检测仪器
酶标仪
检测时长
2h
储存与运输
储存条件
2-8°C储存
运输条件
低温运输
稳定性与储存
各组分在2-8℃储存条件下保质期1年。
组分与储存
E1455939 | Components | 5 Plates | 20 Plates | Storage | Quantity Per Plate |
E1455939A | 包被液 | 55mL | 210mL | 2-8℃ | 10mL |
E1455939B | 封闭液 | 110mL | 410mL | 2-8℃ | 20mL |
E1455939C | 洗涤液(10x) | 200mL | 2*400mL | 2-8℃ | 40mL |
E1455939D | 显色液 A | 30mL | 110mL | 2-8℃ | 5mL |
E1455939E | 显色液 B | 25mL | 85mL | 2-8℃ | 4mL |
E1455939F | 显色液 C | 10mL | 25mL | 2-8℃. Store in the dark | 1mL |
E1455939G | 终止液 | 50mL | 210mL | 2-8℃ | 10 mL |
注：每个Plate推荐使用量为一块96孔酶标板
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