Eotaxin (CCL11) Non-Human Primate ProcartaPlex™ Simplex Kit

Invitrogen™

Eotaxin (CCL11) Non-Human Primate ProcartaPlex™ Simplex Kit

The Non-Human Primate Eotaxin (CCL11) Simplex ProcartaPlex Kit measures Eotaxin (CCL11) protein and is designed to be combinable with other了解更多信息

货号	数量
EPX01B-42120-901	96 次测试

货号 EPX01B-42120-901

价格（CNY)

96 次测试

The Non-Human Primate Eotaxin (CCL11) Simplex ProcartaPlex Kit measures Eotaxin (CCL11) protein and is designed to be combinable with other Simplex kits so that you can create your own multiplex panel that utilizes Luminex xMAP technology for protein detection/quantitation. When combining multiple Simplex kits (i.e., when you are not using a pre-configured Multiplex Panel), only one buffer kit (sold separately) is needed for each assay plate regardless of plex size.

About ProcartaPlex Assays for the Luminex Platform

ProcartaPlex immunoassays are based on the principles of a sandwich ELISA, using two highly specific antibodies binding to different epitopes of one protein to quantitate all protein targets simultaneously using a Luminex instrument. ProcartaPlex multiplex assays require as little as 25 μL of plasma or serum, or 50 μL of cell culture supernatant, and just four hours to obtain analyzed results.

More results per sample—measure multiple protein targets simultaneously in a single 25–50 μL sample
Well-established Luminex technology—highly referenced multiplexing platform for protein detection and quantitation

ProcartaPlex assays utilize Luminex xMAP (multianalyte profiling) technology for the simultaneous detection and quantitation of up to 80 protein targets in a single 25–50 μL sample from plasma, serum, cell culture supernatants, and other bodily fluids.

The Luminex beads in the ProcartaPlex assay are internally dyed with precise proportions of red and infrared fluorophores to create spectrally unique signatures that can be identified by the Luminex xMAP detection systems (e.g. Luminex 200, FLEXMAP 3D, and MAGPIX). Similar to a sandwich ELISA, the ProcartaPlex assay uses matched antibody pairs to identify the protein of interest. In a multiplexed assay, each spectrally unique bead is labeled with antibodies specific for a single target protein, and bound proteins are identified with biotinylated antibodies and streptavidin–R-phycoerythrin (RPE). The conjugation of protein-specific antibodies to a distinct bead allows for analysis of multiple targets in a single well.

The most significant difference between a ProcartaPlex assay and ELISA is that the capture antibody in the ProcartaPlex assay is conjugated to a bead and not adsorbed to the microplate well, so the ProcartaPlex assay reagents are free-floating in the solution. For detection, the Luminex 200 instrument, for example, contains two lasers, one to distinguish the spectral signature of each bead and the second to quantify the amount of RPE fluorescence, which is proportional to the amount of protein present in the sample. ProcartaPlex multiplex assays can profile more target proteins using significantly less sample in the same time that it takes to perform a traditional sandwich ELISA.

ProcartaPlex multiplex panels are available in multiple formats across six species (human, mouse, rat, nonhuman primate, porcine, and canine).

测定范围As determined for Lot 1: 7.56 to 31,000 pg/mL

检测灵敏度根据批次 1 测定：1.4 pg/mL

磁珠类型Eotaxin (CCL11) [33]

适用于（设备）Luminex™ 仪器

产品规格Simplex 试剂盒

基因C-C 基序趋化因子配体 11

基因别名Eotaxin, SCYA11

基因 ID (Entrez)574218

基因符号CCL11

产品线ProcartaPlex

蛋白质C-C motif chemokine ligand 11

样品类型Serum, Plasma, Cell Culture Supernatants

样品体积Serum, Plasma: 25 μL, CCS: 50 μL

运输条件湿冰

UniProt IDQ8HXZ5

可组合性可结合使用

产品类型Simplex 试剂盒

数量96 次测试

研究领域Immunology, Oncology, Neurobiology, Toxicology, Chemokines

种属Non-Human Primate

Unit SizeEach

内容与储存

• 1 vial Capture Beads (50X)
• 1 vial biotinylated Detection Antibody (50X)
• 2 vials NHP Standard Mix A (lyophilized)

Store at 2°C to 8°C.

常见问题解答 (FAQ)

如何处理用于Luminex检测平台的细胞裂解物（提取细胞蛋白质）？

下述方案已经应用于多种人类和小鼠细胞系。您应该为您自己的应用优化细胞提取操作。

细胞裂解操作

非贴壁细胞：
低速离心以沉淀细胞。从沉淀中移去培养基，用冰冷的PBS洗涤两次。去除PBS，将细胞沉淀重悬于细胞裂解缓冲液中（推荐细胞裂解液浓度为2-5 mg/mL）。在冰上孵育15分钟，偶尔涡旋。将裂解物转移至微量离心管中，并在2-8℃下以14000rpm离心10分钟。将澄清的裂解液等分至干净的微量离心管中，并测定总蛋白浓度。

贴壁细胞：
从细胞中去除培养基，用冰冷的PBS洗涤两次。去除PBS，加入细胞裂解缓冲液（推荐细胞裂解液浓度为2-5μg/mL），并在冰上孵育15分钟。收集细胞裂解液，并将其转移至微量离心管中。在2-8℃下以14000rpm离心10分钟。将澄清的裂解液等分至干净的微量离心管中，并测定总蛋白浓度。裂解液应在-80°C下冷冻储存，或在收集后立即进行分析。冷冻样品避免多次反复冻融。分析前，将样品完全解冻、充分混合并通过离心（14000rpm，10分钟）来澄清，以防止堵塞过滤板。

推荐的细胞裂解缓冲液：

NP40细胞裂解缓冲液（货号FNN0021）
注意：使用NP40裂解缓冲液制备的裂解液必须在检测之前稀释至少5倍。
或者使用下述配方进行配置：50 mM Tris，pH 7.4，50 mM NaF，260 mM NaCl，1 mM Na3VO4，5 mM EDTA，0.02% NaN3，1% Nonidet P40

NP40细胞裂解缓冲液（不含蛋白酶抑制剂混合物和PMSF）在2-8°C下的稳定期为2-3周，或在-20°C下分装保存6个月。

使用前在NP40细胞裂解缓冲液中新鲜添加：
•1 mM PMSF（DMSO中的0.3 M原液）
•蛋白酶抑制剂混合物（Sigma，货号P-2714）

替代细胞提取缓冲液

细胞提取缓冲液（货号FNN0011）
注意：使用细胞提取缓冲液制备的裂解液必须在检测之前稀释至少10倍。

或者

10 mM Tris，pH 7.4
2 mM Na3VO4
100 mM NaCl
1% Triton X-100
1 mM EDTA
10% 甘油
1 mM EGTA
0.1% SDS
1 mM NaF
0.5% 脱氧胆酸盐
20 mM Na4P2O7

细胞提取缓冲液（不含蛋白酶抑制剂混合物和PMSF）在2-8°C下的稳定期为2-3周，或在-20°C下等分保存6个月。

使用前在细胞提取缓冲液中新鲜添加：
•1 mM PMSF（DMSO中的0.3 M原液）
•蛋白酶抑制剂混合物（Sigma，货号P-2714）

如何处理在Luminex检测平台上使用的组织匀浆样本？

我们建议遵循下面提供的的方案，这是基于组织提取试剂I（货号FNN0071）开发并显示了ELISA和Luminex技术之间的良好相关性。该方法已应用于多种组织类型。但是，我们建议您针对所使用的每种组织样品进行优化。可以使用类似的提取试剂/裂解缓冲液。

1.在使用前将蛋白酶抑制剂添加到组织提取试剂中。
2.称量组织样品。
3.每1克组织加入10 mL组织提取试剂。
4.匀化组织。
5.以10000rpm离心样品5分钟以沉淀组织碎屑。
6.收集上清液。按照试剂盒中提供的操作步骤进行适当的稀释；这是为了防止/最小化提取液或裂解缓冲液中存在的去垢剂对抗体 - 抗原结合的潜在抑制。通常，组织匀浆或细胞裂解物（取决于所用的裂解缓冲液）需要稀释5至10倍，以将去垢剂浓度降低至≤0.01％。然而，基于目标细胞因子/蛋白质的浓度，不同的试剂盒/样品可能需要用检测稀释液或标准品稀释液进一步稀释。

注意：如果要储存样品，请将其等分并冷冻于-80°C。避免多次反复冻融。

Do you offer a Mac-compatible version of the ProcartaPlex Analyst Software?

Unfortunately, we do not currently offer a Mac-compatible version of the ProcartaPlex Analyst Software. ProcartaPlex Analyst Software is compatible with PC, or xPONENT Software is available on Luminex instruments to analyze your data.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I am getting poor recovery with my ProcartaPlex multiplex immunoassay. What can I do?

Here are possible causes and solutions:
- The wrong cell culture media may have been used to prepare the standards. We recommend using the same cell culture media that was used to culture the cells.
- The samples and antigen standards may not have been stored on ice. We recommend preparing the samples and standards on ice before setting up the assay.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I am seeing low signal or sensitivity in my ProcartaPlex multiplex immunoassay. What should I do?

Standards may not have been reconstituted and diluted correctly. We recommend preparing fresh antigen standards following the instructions provided in the corresponding ProcartaPlex user guide.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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