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电话: 400-6721-600 Mouse IgE ELISA Kit This package insert must be read entirely before using this product. For proper performance, use the insert provided with each individual kit received. Catalog Number EK2751 C

48 T EK2752 C

96 T EK2753 C

2 *96 T EK2754 C

5 *96 T EK2755 C

10 *96 T For the quantitative determination of mouse Immunoglobulin E (IgE) concentration in cell culture supernates, serum and plasma. For research use only. Not for use in diagnostic procedures. Mouse IgE ELISA Kit 08/2016

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电话: 400-6721-600 Mouse IgE ELISA Kit Table of Contents Introduction Description

1 Principle of the Assay

1 Limitations of the Procedure

1 General Information Materials Provided

3 Other Supplies required

3 Storage

5 Precaution

7 Technical Hints

9 Assay Protocol Sample Collection and Storage

9 Sample Preparation

11 Reagent Preparation 11,13 Assay Procedure

15 Analysis Calculation of Results

17 Typical Data

17 Sensitivity

19 Precision

19 Calibration

19 Sample Values

21 Assay Procedure Summary

23 目录 产品介绍 背景介绍

2 检测原理

2 试剂盒检测的局限

2 基本信息 试剂盒提供的材料

4 未提供的材料设备

4 贮存

6 注意事项

8 技术要点

10 检测步骤 样本的收集和贮存

10 试剂准备 12,

14 检测步骤

16 分析 结果计算

18 典型数据

18 灵敏度

20 精密度

20 校准

20 样本值

22 检测步骤概要

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电话: 400-6721-600 Mouse IgE ELISA Kit DESCREPTION Immunoglobulin E (IgE) is a kind of antibody that has only been found in mammals. IgE'

s main function is immunity to parasites such as helminths, Schistosoma mansoni, Trichinella spiralis, and Fasciola hepatica. IgE also has an essential role in type I hypersensitivity, which manifests various allergic diseases, such as allergic asthma, most types of sinusitis, allergic rhinitis, food allergies, and specific types of chronic urticaria and atopic dermatitis. IgE also plays a pivotal role in responses to allergens, such as: anaphylactic drugs, bee stings, and antigen preparations used in desensitization immunotherapy. Although IgE is typically the least abundant isotype ― blood serum IgE levels in a normal individual are only 0.05% of the Ig concentration ― it is capable of triggering the most powerful inflammatory reactions. Atopic individuals can have up to

10 times the normal level of IgE in their blood. IgE is known to be elevated in various autoimmune disorders such as Lupus (SLE), Rheumatoid Arthritis (RA) and psoriasis, and is theorized to be of pathogenetic importance in RAand SLE by eliciting a hypersensitivity reaction. PRINCIPLE OF THE ASSAY This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for mouse IgE has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and IgE present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-linked detect antibody specific for IgE is added to the wells. Following a wash to remove any unbound antibody-biotin reagent, streptavidin-HRP is added. After washing, substrate solution is added to the wells and color develops in proportion to the amount of IgE bound in the initial step. The color development is stopped and the intensity of the color is measured. LIMITATIONS OF THE KIT FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. The kit should not be used beyond the expiration date on the kit label. Do not mix or substitute reagents with those from other lots or sources. If samples generate values higher than the highest standard, dilute the samples with Assay Buffer and repeat the assay. If cell culture supernate samples require large dilution, perform an intermediate dilution in culture medium. Any variation in standard dilution, operator, pipetting technique, washing techniques, incubation time or temperature, and kit age can cause variation in binding. This assay is designed to eliminate interference by factors present in biological samples. Until all factors have been tested in the ELISA immunoassay, the possibility of interference cannot be excluded. 背景介绍 免疫球蛋白 E (IgE)是仅存在于哺乳动物中的一类抗体. IgE 的主要功能是对寄生虫产生免疫, 如蠕虫、 曼氏血吸虫、 旋毛虫和肝片吸虫. IgE 在I型超敏反应中具有关键性作用, 如过敏性哮喘、 大部分鼻窦炎、变应性鼻炎、食物过敏和慢性荨麻疹和特应性皮炎等.IgE 在对变应原产生应答中 也具有重要作用,如过敏性药物、蜂蜇伤、脱敏免疫治疗中用到的抗原.虽然 IgE 含量最少(正常 人血清中的 IgE 含量仅占所有 Ig 类的 0.05 %), 但它能触发最强烈的炎症反应. 遗传性过敏症患者 血液中的 IgE 含量高达正常人的

10 倍.在多种自身免疫疾疾病如系统性红斑狼疮(SLE)、类风湿 性关节炎(RA)和银屑病中,IgE 水平均有上升,推测其通过诱导超敏反应对 RA 和SLE 具有重要 的致病作用. 检测原理 本试剂盒采用双抗体夹心酶联免疫吸附检测技术.特异性抗小鼠 IgE 单克隆抗体预包被在高 亲和力的酶标板上.酶标板孔中加入标准品和待测样本,经过孵育,样本中存在的 IgE 与固相抗 体结合.洗涤去除未结合的物质后,加入生物素化的检测抗体孵育.洗涤去除未结合的生物素化 的抗体, 加入辣根过氧化物酶标记的链霉亲和素 (Streptavidin-HRP). 洗涤后, 加入显色底物 TMB, 避光显色.颜色反应的深浅与样本中 IgE 的浓度成正比.加入终止液终止反应,在450 nm 波长(参 考波长

570 -

630 nm)测定吸光度值. 试剂盒检测的局限 1. 本试剂盒用于科学研究,非诊断试剂,不能用于临床诊断. 2. 请在本试剂盒标记的有效期内使用. 3. 试剂盒的试剂不能与其他批号的试剂或其他来源的试剂混合使用. 4. 如果样本值高于标准曲线最高浓度,请用检测缓冲液稀释样本,并重新检测.如果细胞培养上 清样本需要较大的稀释倍数,请用细胞培养基进行适度稀释. 5. 任何标准品稀释、操作人员、移液技术、洗涤技术、孵育温度、试剂盒保存时间的改变,都将 影响结合反应. 6. 本试剂盒在设计上去除或降低了生物学样本中的一些内源性干扰因素,并非所有可能的影响因 素都已经去除.

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电话: 400-6721-600 Mouse IgE ELISA Kit MATERIALS PROVIDED 1. IgE Microplate:

96 well polystyrene microplate (12 strips of

8 wells) coated with a monoclonal antibody against mouse IgE. 2. IgE Standard: Recombinant mouse IgE in a buffered protein base with preservatives;

lyophilized. 3. IgE Detect Antibody: Biotin-conjugate anti-mouse IgE detect antibody;

100*liquid. 4. Streptavidin-HRP: 100*liquid. 5. Assay Buffer (10* ): PBS with 0.5 % Tween-20 and

5 % BSA. 6. Substrate: TMB (tetramethyl-benzidine). 7. Stop Solution: 0.18M sulfuric acid. 8. Wash Buffer (20* ): PBS with

1 % Tween-20. 9. Plate Covers. All reagents should be stored at

2 - 8℃. OTHER SUPPLIES REQUIRED 1. Microplate reader capable of measuring absorbance at

450 nm, with correction wavelength set at

570 nm or

630 nm. 2. Pipettes and pipette tips. 3.50 μl to

300 μl adjustable multichannel micropipette with disposable tips. 4. Multichannel micropipette reservoir. 5. Beakers, flasks, cylinders necessary for preparation of reagents. 6. Deionized or distilled water. 7. Polypropylene test tubes for dilution. 试剂盒提供的材料 未提供的材料设备 1. 能够检测

450 nm 吸光度的酶标仪,参考波长

570 nm 或630 nm. 2. 移液器及枪头 3.

50 -

300 μl 可调多道移液器及一次性枪头 4. 多道移液器加样槽 5. 准备试剂用的试管、离心管、量筒等 6. 蒸馏水或去离子水 7. 稀释用聚丙烯试管 组分 编号 EK2751 EK2752 EK2753 EK2754 EK2755 预包被酶标板 EK275P 48T 96T 192T 480T 960T 标准品 EK275S

1 vial

2 vial

4 vial

10 vial

20 vial 检测抗体 EK275D

1 vial

1 vial

2 vial

5 vial

10 vial 辣根过氧化物酶标 记的链霉亲和素 E0290

1 vial

1 vial

2 vial

5 vial

10 vial 10* 检测缓冲液 E0310

10 ml

10 ml

20 ml

50 ml

100 ml 显色底物 TMB E0230

11 ml

11 ml

22 ml

55 ml

110 ml 终止液 E0300

11 ml

11 ml

22 ml

55 ml

110 ml 20* 洗液 E0281

50 ml

50 ml

100 ml

250 ml

500 ml 封板膜 E0200

3 3

6 15

30 3

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电话: 400-6721-600 Mouse IgE ELISA Kit STORAGE Store kit reagents between

2 and 8℃. Immediately after use remaining reagents should be returned to cold storage (2 to 8℃). Expiry of the kit and reagents is stated on labels. Expiry of the kit components can only be guaranteed if the components are stored properly, and if, in case of repeated use of one component, this reagent is not contaminated by the first handling. Unopened kit Store at

2 - 8℃. Opened/ Reconstituted Reagents Diluted Wash Buffer Diluted Assay Buffer Stop Solution Substrate TMB Detect Antibody Streptavidin-HRP May be stored for up to

1 month at

2 - 8℃ Standard for up to

1 month at ≤ -20℃ in a manual defrost freezer. Discard after use. Microplate Wells Return unused wells to the foil pouch containing the desiccant pack, reseal along entire edge of zip-seal. May be stored for up to

1 month at

2 - 8℃. Provided this is within the expiration date of the kit. 贮存 试剂盒保存于

2 - 8℃,有效期标注于标签上.只有恰当保存的试剂才是有保证的.如果试剂 盒的组分需要再次使用,请确定上一次使用之后试剂没有污染. 未开封试剂盒 贮存于

2 - 8℃.请在有效期内使用. 打开的试剂盒或重组试剂1* 洗液 1* 检测缓冲液 终止液 底物 TMB 检测抗体 辣根过氧化物酶标记的链霉亲和素 在2-8℃,大约可以贮存

1 个月. 标准品 置于-20℃,大约可贮存

1 个月. 使用后丢弃. 预包被酶标板 未使用的板条请放回铝箔袋,封好封口. 在2-8℃,大约可贮存

1 个月.

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