Polystyrene microspheres present a flexible platform for applications in diagnostics and bioseparations. They may be coated with recognition molecules, such as antibodies, antigens, peptides, or nucleic acid probes, and can be loaded with hydrophobic dyes and other compounds. Unmodified polymer spheres also find extensive use as standards for instrument set-up and calibration. Available in diameters ranging from 20nm to 200µm, products exhibit excellent size uniformity.
聚苯乙烯微球为诊断和生物分离应用提供了一个灵活的平台。它们可以涂有识别分子，例如抗体、抗原、肽或核酸探针，并且可以装载疏水性染料和其他化合物。未改性的聚合物球也广泛用作仪器设置和校准的标准。产品的直径范围从 20nm 到 200µm，具有出色的尺寸均匀性。
Plain polystyrene microspheres are ideal for protein adsorption, and have been utilized in a range of diagnostic tests and assays. Surface modified microspheres are available with carboxyl or primary amine groups for covalent ligand attachment. Affinity binding systems offer simple and efficient ligand attachment. Coatings of Fc binding proteins are able to orient antibodies for optimal activity, and streptavidin offers extremely stable attachment of biotinylated molecules, such as proteins, peptides, and oligonucleotides.
普通聚苯乙烯微球是蛋白质吸附的理想选择，并已用于一系列诊断测试和分析。表面改性的微球具有羧基或伯胺基，用于共价配体连接。亲和结合系统提供简单有效的配体连接。Fc 结合蛋白涂层能够使抗体定向以获得最佳活性，而链霉亲和素可提供极其稳定的生物素化分子（如蛋白质、肽和寡核苷酸）附着。
QuantumPlex™ SP Streptavidin Product Data Sheet 214
Description The QuantumPlex™ kit is designed to be used as a multiplexing platform, allowing for the efficient, qualitative flow cytometric analysis of a sample for multiple analytes, or the high throughput screening of multiple samples. QuantumPlex™ SP is a single population of microspheres sized 4.4µm (Catalog Code 214) or 5.5µm (Catalog Code 217). The beads are internally dyed with Starfire Red™ fluorescent dye (fluorescent in FL3). The beads have a uniform streptavidin (SA) surface. The SA surface allows for the easy conjugation of analytes or analyte-specific antibodies to the surface of each bead. The beads may then be incubated with a sample and washed before a fluorescently-tagged reported antibody is added. After a second wash and resuspension, the beads may be analyzed with a flow cytometer to determine the presence or absence of the assayed analyte. Conjugation techniques optimized for the single population of QuantumPlex™ SP beads may be easily applied to the 5-bead QuantumPlex™ multiplexing bead array.
描述
QuantumPlex™ 试剂盒设计用作多路复用平台，允许对样品进行高效、定性的流式细胞仪分析以了解多种分析物，或对多种样品进行高通量筛选。 QuantumPlex™ SP 是一组大小为 4.4µm（目录代码 214）或 5.5µm（目录代码 217）的微球。珠子内部用 Starfire Red™ 荧光染料（FL3 中的荧光）染色。珠子具有均匀的链霉亲和素 (SA) 表面。 SA 表面允许将分析物或分析物特异性抗体轻松结合到每个珠子的表面。然后可以将珠子与样品一起温育并在添加荧光标记的报告抗体之前进行洗涤。在第二次洗涤和再悬浮之后，可以用流式细胞仪分析珠子以确定被分析的分析物的存在或不存在。针对单个 QuantumPlex™ SP 珠子群优化的偶联技术可轻松应用于 5 珠 QuantumPlex™ 多路复用珠子阵列。
特征
Mean Diameter: 4.4µm (Catalog Code 214) or 5.5µm (Catalog
Code 217)
Particle Concentration: 1 x 10^6microspheres/mL
Material
提供的材料
• QuantumPlex™ SP microspheres: bottled individually in 1mL or
3mL aliquots
• Storage Buffer: 0.1% BSA, 0.05% Tween® 20, and 10mM EDTA
所需材料
• Biotinylated analyte or antibody specific to the analyte(s) of
interest
• Fluorescently-labeled reporter antibody (fluorescent in FL1 or FL2)
Procedure
Researchers are advised to optimize the use of particles in any application.QuantumPlex™ SP allows for flexibility in designing individual experiments.
The preparation procedure outlines the conjugation of a single biotinylated antibody to the QuantumPlex™ bead. In doing so, an assay may be produced which is capable of testing a single sample for a single analyte.The user may choose instead to conjugate multiple antibodies of different
specificities to the beads, producing an assay ideal for screening a sample for multiple analytes in a single test. The user may further choose to conjugate antigen to beads, yielding an assay capable of testing for the presence of a specific antibody. The specific application is to be determined by the user. The following outline serves as a guide, and may be modified to reflect the user’s specific application. For a more detailed coupling procedure, see TechNote 101, ProActive® Microspheres.
建议研究人员在任何应用中优化粒子的使用。
QuantumPlex™ SP 允许灵活设计单个实验。
制备程序概述了单个生物素化的缀合QuantumPlex™ 微珠的抗体。 在这样做的时候，一个化验可能是生产的能够测试单一样品的单一分析物。
用户可以选择缀合多种不同的抗体对珠子的特异性，产生筛选样品的理想化验在一次测试中检测多种分析物。 用户可以进一步选择将抗原与珠子结合，产生能够测试
存在特异性抗体。 具体申请待定由用户。 以下大纲作为指导，可能会被修改反映用户的具体应用。 更详细的耦合步骤，参见技术说明 101，ProActive® 微球。
Preparation of Microspheres
1. Vortex the bottle prior to use to ensure uniform suspension of the
beads.
2. Immediately remove 10µL of solution to be labeled with ligand. Note:The 10µL volume reflects the amount needed to conduct one test using the given bead population. For ease of use, the entire 1mL or 3mL may be labeled all at once, and then stored for use with each test.
3. Using an excess of biotinylated antigen or antibody, conjugate the antigen or antibody to the bead using established protocols. Note:One approach is to simply incubate the beads and the biotinylated antigen or antibody together for 30 minutes. See also TechNote 101, ProActive® Microspheres, for sample conjugation protocols.
4. Wash the beads to remove any unbound antibody.
5. Resuspend beads in ~100µL buffer.
1. 使用前涡旋瓶子以确保均匀悬浮珠子。
2. 立即取出 10µL 待用配体标记的溶液。 笔记：10µL 体积反映了使用给定的珠子数量。 为方便使用，整个 1mL 或 3mL 可一次全部贴上标签，然后储存起来以供每次测试使用。
3. 使用过量的生物素化抗原或抗体，将使用已建立的协议对珠子进行抗原或抗体。 笔记：一种方法是简单地孵育珠子和生物素化的抗原或抗体一起 30 分钟。 另见技术说明 101，ProActive® 微球，用于样品偶联方案。
4. 清洗磁珠以去除任何未结合的抗体。
5. 在~100µL 缓冲液中重悬珠子。
Testing Samples
1. Incubate prepared beads with 100µL sample(s) for 30 minutes. (The volume of sample used may be adapted to the specific application.)
2. Wash beads to remove nonspecifically bound analyte. Repeat the wash step.
3. Incubate the beads with 20µL of the appropriate fluorescently-labeled antibody for 30 minutes.
4. Wash beads to remove nonspecifically bound antibody. Repeat the wash step.
5. Acquire data events using a flow cytometer
1. 将制备好的珠子与 100 µL 样品一起孵育 30 分钟。 （使用的样品量可根据具体应用进行调整。）
2. 清洗珠子以去除非特异性结合的分析物。 重复洗涤步骤。
3. 将微珠与 20µL 适当的荧光标记抗体一起孵育 30 分钟。
4. 清洗磁珠以去除非特异性结合的抗体。 重复洗涤步骤。
5. 使用流式细胞仪获取数据事件
Data Analysis
1. Gate on the single population(s) on a Forward Scatter vs. Side Scatter plot. (Figure 4)
2. Using the FL1 and / or FL2 channels (depending on the reporter antibodies used), determine whether or not any bead populations tested “positive” for the analyte. (Figure 5) Note: A positive bead will produce a fluorescent peak in the FL1 or FL2 channel. The minimum fluorescence intensity needed to be considered “positive” is based on the Relative Channel Value (RCV) of the peak. It is up to the investigator to determine what threshold RCV value will constitute a “positive” result.
3. The intensity of the Starfire Red™ dye contained in the bead is used to differentiate the bead from others in the 5-bead QuantumPlex™ kit. (Figure 6) When using only the QuantumPlex™ SP beads, the use of “back-gating” on the red (FL3) signal may be performed to rule out debris and validate your correct identification of the beads.
1. 前向散射与侧向散射上单个种群的门阴谋。 （图4）
2. 使用 FL1 和/或 FL2 通道（取决于记者使用的抗体），确定是否测试了任何珠群分析物为“阳性”。 （图 5） 注意：一个阳性珠子会产生一个
FL1 或 FL2 通道中的荧光峰。 最小荧光需要被视为“积极”的强度是基于相对峰值的通道值 (RCV)。 由调查人员决定确定哪个阈值 RCV 值将构成“阳性”结果。
3. 使用珠子中包含的 Starfire Red™ 染料的强度将 5 珠 QuantumPlex™ 中的珠子与其他珠子区分开来成套工具。 （图 6）仅使用 QuantumPlex™ SP 微珠时，使用
可以对红色 (FL3) 信号执行“反向选通”以排除碎片并验证您对珠子的正确识别。
Trademarks and Registered Trademarks
1. QuantumPlex™, Starfire Red™, and ProActive® are trademarks or
registered trademarks of Bangs Laboratories, Inc.
2. Tween® is a registered trademark of ICI Americas, Inc.
Storage and Stability
Store at 2-8˚C. Freezing may result in irreversible aggregation and loss of binding activity. QuantumPlex™ SP beads are stable for 12 months from date of purchase, provided the product is handled in accordance with the manufacturer’s recommendations. The beads should be kept in the bottle in which they are shipped. Do not expose the beads to intense light sources for extended periods of time.
储存于 2-8°C。 冷冻可能导致不可逆的聚集和损失结合活性。 QuantumPlex™ SP 微珠可稳定 12 个月，从购买日期，提供的产品是按照制造商的建议。 珠子应保存在瓶中他们被运送。 不要将珠子暴露在强光源下延长的时间。
Safety
This particle suspension contains sodium azide. Sodium azide may react with lead and copper plumbing to form explosive metal azides. Upon disposal of material, flush with a large volume of water to prevent azide accumulation.Please consult the Material Safety Data Sheet for more information.These products are for research use only and are not intended for use in humans or for in vitro diagnostic use.
这种颗粒悬浮液含有叠氮化钠。 叠氮化钠可能与铅和铜管道形成爆炸性金属叠氮化物。 处置后材料，用大量水冲洗以防止叠氮化物堆积。请查阅材料安全数据表了解更多信息。这些产品仅供研究使用，不适用于用于人类或体外诊断用途。
Ordering Information
Cat. Code Description Sizes
214 QuantumPlex™ SP Streptavidin 4.4µm 1mL or 3mL
217 QuantumPlex™ SP Streptavidin 5.5µm 1mL or 3mL