PDF《网址：www.liankebio.com 客服电话：》

网址: www.

liankebio.com 客服

电话: 400-6721-600 Human Ephrin-A1/EFNA1 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 EK11891 C

48 T EK11892 C

96 T EK11893 C

2 *96 T EK11894 C

5 *96 T EK11895 C

10 *96 T For the quantitative determination of human EPH-related Receptor Tyrosine Kinase Ligand

1 (Ephrin-A1/EFNA1) concentration in cell culture supernates, serum and plasma. For research use only. Not for use in diagnostic procedures. 08/2016 Human Ephrin-A1/EFNA1 ELISA Kit

网址: www.liankebio.com 客服

电话: 400-6721-600 Human Ephrin-A1/EFNA1 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 Reagent Preparation 11,

13 Assay Procedure

15 Analysis Calculation of Results

17 Typical Data

17 Sensitivity

19 Precision

19 Recovery

21 Linearity

21 Calibration

21 Sample Values

23 Specificity

23 Assay Procedure Summary

25 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 回收率

22 稀释线性

22 校准

22 样本值

24 特异性

24 检测步骤概要

26

网址: www.liankebio.com 客服

电话: 400-6721-600 Human Ephrin-A1/EFNA1 ELISA Kit DESCREPTION Ephrin-A1, also known as EFNA1, is a protein that in humans is encoded by the EFNA1 gene. This gene encodes a member of the ephrin (Eph) family. The ephrins and Eph-related receptors comprise the largest subfamily of receptor protein-tyrosine kinases and have been implicated in mediating developmental events, especially in the nervous system and in erythropoiesis. Based on their structures and sequence relationships, ephrins are divided into the ephrin-A (EFNA) class and the ephrin-B (EFNB) class. Ephrin-A ligands bind EphA receptors and are anchored to the plasma membrane via a glycosylphosphatidylinositol (GPI) linkage, whereas ephrin-B ligands bind EphB receptors and are anchored via a transmembrane domain. Ephrin-A1 and one of its receptor EphA2 were expressed in xenograft endothelial cells and also tumor cells and play a role in human cancers, at least in part by influencing tumor neovascularization. PRINCIPLE OF THE ASSAY This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for human Ephrin-A1 has been pre-coated onto a microplate. Standard, samples and HRP-linked detect antibody specific for Ephrin-A1 are pipetted into the wells and Ephrin-A1 present is bound by the immobilized antibody and detect antibody following incubation. After washing, substrate solution is added to the wells and color develops in proportion to the amount of Ephrin-A1 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 Ephrin-A1,又名 EFNA1,在人类中由 EFNA1 基因编码,该基因编码 Eph 家族成员.ephrins 和Eph 相关受体组成了最大的受体酪氨酸激酶亚家族,参与调节神经和造血系统的发展.根据它 们的结构和序列关系,ephrins 可分为 ephrin-A(EFNA)类和 ephrin-B(EFNB)类.Ephrin-A 配体结合 EphA 受体,通过糖基磷脂酰肌醇(GPI)连接锚定到质膜上.而ephrin-B 配体结合 EphB 受体,通过 跨膜结构域锚定到质膜上.Ephrin-A1 及它的其中之一受体 EphA2 在异种移植的内皮细胞和肿瘤 细胞中表达,在某种程度上可通过影响肿瘤新血管形成,从而在人类癌症中发挥作用. 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. 特异性抗人 Ephrin-A1 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、待测样本和辣根过氧化物酶标记的检测抗体,经 过孵育,样本中存在的 Ephrin-A1 与固相抗体和检测抗体结合.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 Ephrin-A1 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.

2 1

网址: www.liankebio.com 客服

电话: 400-6721-600 Human Ephrin-A1/EFNA1 ELISA Kit MATERIALS PROVIDED 1. Ephrin-A1 Microplate:

96 well polystyrene microplate (12 strips of

8 wells) coated with a monoclonal antibody against human Ephrin-A1. 2. Ephrin-A1 Standard: Recombinant human Ephrin-A1 in a buffered protein base with preservatives;

lyophilized. 3. Ephrin-A1 Detect Antibody: HRP-conjugate anti-human Ephrin-A1 detect antibody;

100*liquid. 4. Standard Diluent: In some, very rare cases, an insoluble precipitate of stabilizing protein has been seen in the Standard Diluent vial. This precipitate does not interfere in any way with the performance of the test and can thus be ignored. 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 EK11891 EK11892 EK11893 EK11894 EK11895 预包被酶标板 EK1189P 48T 96T 192T 480T 960T 标准品 EK1189S

1 vial

2 vial

4 vial

10 vial

20 vial 辣根过氧化物酶 标记的检测抗体 EK1189D

1 vial

1 vial

2 vial

5 vial

10 vial 标准品稀释液 EKSP05

5 ml

5 ml

10 ml

25 ml

50 ml 10* 检测缓冲液 E0310

5 ml

5 ml

10 ml

25 ml

50 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

4

网址: www.liankebio.com 客服

电话: 400-6721-600 Human Ephrin-A1/EFNA1 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 Standard Diluent Substrate TMB Detect Antibody 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 标准品稀释液 底物 TMB 检测抗体 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.

5 6

网址: www.liankebio.com 客服

电话: 400-6721-600 Human Ephrin-A1/EFNA1 ELISA Kit PRECAUTION 1. All chemicals should be considered as potentially hazardous. 2. We therefore recommend that this product is handled only by those persons who have been trained in laboratory techniques and that it is used in accordance with the principles of good laboratory practice. Wear suitable protective clothing such as laboratory overalls, safety glasses and gloves. 3. Care should be taken to avoid contact with skin or eyes. In the case of contact with skin or eyes wash immediately with water. See material safety data sheet(s) and/or safety statement(s) for specific advice. 4. The Stop Solution provided with this kit is an acid solution. Wear eyes, hand, face, and clothing protection when using this material. 5. Reagents are intended for research use only and are not for use in diagnostic or therapeutic procedures. 6. Do not mix or substitute reagents with those from other lots or other sources. 7. Do not use kit reagents beyond expiration date on label. 8. Do not expose kit reagents to strong light during storage or incubation. 9. Do not eat or smoke in areas where kit reagents or samples are handled. 10. Avoid contact of skin or mucous membranes with kit reagents or specimens. 11. Rubber or disposable latex gloves should be worn while handling kit reagents or specimens. 12. Avoid contact of substrate solution with oxidizing agents and metal. 13. Avoid splashing or generation of aerosols. 14.In order to avoid microbial contamination or cross- contamination of reagents or specimens which may invalidate the test use disposable pipette tips and/or pipettes. ........