Dynabeads™ Epithelial Enrich
Dynabeads™ Epithelial Enrich
Invitrogen™

Dynabeads™ Epithelial Enrich

Dynabeads Epithelial Enrich contains 4.5-μm superparamagnetic beads coupled to a monoclonal mouse anti-human EpCAM antibody and is intended for isolation of circulating epithelial tumor cells (CTCs) from liquid biopsy samples.
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货号数量
161025 mL
16103D2 mL
货号 16102
价格(CNY)
17,536.00
飞享价
Ends: 31-Dec-2025
24,261.00
共减 6,725.00 (28%)
Each
添加至购物车
数量:
5 mL
价格(CNY)
17,536.00
飞享价
Ends: 31-Dec-2025
24,261.00
共减 6,725.00 (28%)
Each
添加至购物车

Dynabeads Epithelial Enrich consist of 4.5-μm superparamagnetic beads coupled to a monoclonal mouse anti-human EpCAM antibody and are intended for isolation of circulating epithelial tumor cells (CTCs) from liquid biopsy samples. The sensitive isolation method provided by Dynabeads Epithelial Enrich is necessary given the low number of CTCs in liquid biopsy samples. The isolated CTCs remain attached to the beads throughout the process.

For downstream molecular or proteomic analysis, the cells can be directly lysed while still attached to the beads. RT-PCR can be performed for amplification and quantification of target nucleic acids. CTC isolation and any subsequent bead-based proteomic or molecular applications can be performed manually or automated using the KingFisher Apex purification system.

Benefits of Dynabeads Epithelial Enrich include:

  • High sensitivity and specificity isolation of CTCs from liquid biopsy samples
  • Fast and efficient isolation of viable CTCs
  • Automation user guide and script available for high-throughput enrichment using the KingFisher Apex instrument

Dynabeads-based positive isolation of circulating epithelial tumor cells

Dynabeads Epithelial Enrich are added to the sample under continuous mixing for 30 minutes to optimize the binding of the Dynabeads to the target CTCs. By placing the sample in a DynaMag magnet, the bead-bound epithelial cells are separated from the rest of the sample in l 2 minutes. The supernatant is discarded, and the bead-bound cells are washed 2 to 3 times to obtain optimal purity. The cells can be lysed while still attached to the beads and the supernatant transferred to a new tube for cell culture or downstream molecular or proteomic analysis. Gene expression analysis without pre-enrichment of RNA can easily be performed using the TaqMan Cells-to-CT Express Kit. Starting samples can be whole blood, buffy coat, bone marrow, or peripheral blood mononuclear cells (PBMCs).

High throughput positive isolation of CTCs on KingFisher Apex instrument

The entire workflow can be automated using the KingFisher Apex purification system with 96-deep-well plates. The high-throughput recovery of bead-bound CTCs can be achieved in about 50 minutes after the sample and reagents are loaded into the plates. The isolated CTCs are viable and suitable for culture and downstream molecular or proteomic analysis. Benefits of using automated CTC isolation include:

  • Reduced hands-on time
  • High reproducibility due to fewer manual handling errors
  • Increased throughput with high purity of viable CTCs.

Alternative CTC enrichment methods

Positive isolation with release: If it is preferred to positively isolate CTCs for immunological applications such as flow cytometry, we recommend using Dynabeads CELLection Epithelial Enrich (Cat. No. 16203) where the beads are released from the cells after the isolation. The bead-free CTCs are pure, viable and can be used in any downstream application. If you are using your own biotinylated antibody, we recommend using Dynabeads MyOne Streptavidin T1.

Negative isolation of untouched CTCs: If it is preferred to isolate untouched CTCs by leukocyte depletion, we recommend using Dynabeads CD45 (Cat. No. 11153D).

Commercial supply

Our manufacturing sites are ISO 13485-certified. Bead characteristics and manufacturing conditions help ensure consistency and batch reproducibility making them well suited for commercial supply. If you are in the process of customizing Dynabeads Epithelial Enrich in a commercial product or service or need a larger bulk volume of this product, please contact us at oemdynal@lifetech.com or read more on our Dynabeads OEM page.

仅供科研使用。不可用于诊断程序。

规格
细胞类型肿瘤细胞、上皮细胞
包括5 mL Dynabeads coated with the monoclonal antibody BerEP4 against the human epithelial antigen, EpCAM
分离技术用于分子应用的去除、阳性分离
池数量处理总计∼4x10^9个细胞
输出可行性>95%
产品线DYNAL™,Dynabeads™
纯度或质量等级研究级
数量5 mL
反应性
样品类型PBMC、血液
运输条件室温
原始材料细胞数量一次试验可分离 2x10^7 PBMC
目标种属
产品类型抗体包被微球
Unit SizeEach
内容与储存
本产品含:5 mL Dynabeads™,包被抗人
上皮抗原 EpCAM 的单克隆抗体 BerEP4。
在 2-8°C 下储存

常见问题解答 (FAQ)

我的Dynabeads磁珠不能很好地吸附到磁力架上,对此你们有什么建议吗?

请查看以下可能原因:

•溶液太粘稠。
•蛋白质间相互作用导致磁珠聚集。

尝试以下建议:
•延长分离时间(将管子留在磁力架上2-5分钟)。
•向裂解液中加入DNase I(约0.01 mg/mL)。
•将结合和/或清洗缓冲液中的Tween20浓度增加至约0.05%。
•向结合和/或清洗缓冲液中加入最多至20 mM 的β-巯基乙醇。

我想要分离较长的双链DNA片段,你们有什么产品可以推荐?

对于小于1 kb的生物素标记DNA,我们推荐使用Dynabeads M270链霉亲和素磁珠和MyOne C1磁珠。对于大于1kb的双链DNA分子,我们推荐Dynabeads KilobaseBINDER试剂盒。KilobaseBINDER试剂包括M-280链霉亲和素偶联的Dynabeads磁珠和一种含有专利的固定活化剂的结合液,可结合较长的生物素化DNA分子以进行分离。请点击以下链接(https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/immobilisation-of-long-biotinylated-dna-fragments.html),查看关于长的生物素化DNA片段分离的更多信息。

我能否使用Dynabeads磁珠分离单链DNA模板?

可以,Dynabeads磁珠可用于分离单链DNA。链霉亲和素Dynabeads磁珠能够以生物素化的DNA片段为靶标,通过使双链DNA变性,从而去除非生物素化链。链霉亲和素偶联的Dynabeads磁珠不会抑制任何酶活性。因此,可以在固相上直接对磁珠结合的DNA进行下一步处理。请点击以下链接(https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/preparing-single-stranded-dna-templates.html),查看关于单链DNA捕获的更多信息。

什么是磁化率?

磁化率能够衡量磁珠向磁力架迁移的速度,其大小取决于铁含量和氧化铁的特性。Dynabeads磁珠的磁化率是指质量磁化率,单位可以是cgs单位/g或m^3/kg(国际单位制)。对于亚铁磁性和铁磁性物质,质量磁化率取决于磁场强度(H),这些物质的磁化强度与H不是线性关系,而是随着场强增加而趋于饱和。因此, Dynabeads磁珠的质量磁化率是在固定条件下由标准操作程序而测定的。我们产品目录中给出的质量磁化率是国际单位制。磁化率由从高斯(cgs、emu)单位向国际单位的转换,是通过“高斯系数(emu/g或cgs/g)x 4π x 10^-3”而实现的。所得单位也被称为合理化质量磁化率,与(国际单位制)无量纲磁化率单位有所区别。通常,质量磁化率可用来衡量在非均匀磁场中影响物体的力(Fz)。测定Dynabeads磁珠的质量磁化率时,首先对样本称重,然后将样本放置于已知强度的磁场中。随后,再次称重得到样本重量(F1),并与关闭磁场时样本的重量(F0)进行对比。使用下述公式计算磁化率:K x 10^–3 = [(F1-F0) x m x 0.335 x 10^6],K表示质量为m的样本的质量磁化率。最后,将磁化率转换为国际单位制。

我如何确定Dynabeads磁珠的偶联效率?

有多种不同的方法可以检测配体与磁珠结合,包括光密度(OD)检测、荧光标记和放射性标记。

对于OD检测,应在配体固定到磁珠上之前检测配体的OD值,并将其与包被后上清液中剩余的配体浓度进行比较。这样可以粗略检测有多少蛋白与磁珠结合。 实验方案: 1.将分光光度计设置到正确的波长。使用偶联缓冲液作为空白组。 2.检测偶联前溶液的吸光值。根据配体的加入量,可能需要进一步稀释以读取吸光值。 3.检测偶联后溶液的吸光值。也可能需要进一步稀释以读取吸光值。 4.计算偶联效率,以“蛋白质摄取量%”表示,如下所示:[(偶联前溶液的吸光值x D) – (偶联后溶液的吸光值x D)] x 100/(偶联前溶液的吸光值 x D),D = 稀释倍数。 对于荧光标记,我们建议对配体结合量进行反向定量,即检测偶联上清液中剩余的配体量(与原始样本对比),而不是直接检测磁珠上的配体量。将标记的配体加入到磁珠中,并检测上清液中剩余多少配体(而不是结合到磁珠上的配体)。通过与开始时加入的总配体量相比,可以计算出结合到磁珠上的配体量。由于Dynabeads磁珠具有自发荧光,因此,我们不推荐直接检测与磁珠结合的配体的荧光,而是推荐这种间接方法。标记物可以是FITC/PE等。有些研究人员也成功使用了直接检测方法(采用流式细胞仪)。 在3种方法中,放射标记的灵敏度最高,但难度最大。该方法涉及到对配体的一部分进行放射性标记。在偶联前,使用示踪剂量的放射性标记的I-125,将其以一定比例与“冷”配体混合。使用闪烁(γ)计数器对磁珠进行检测,并将磁珠的cpm值与标准品对比,得到磁珠上配体的绝对量。 实验方案: 1.取出适量磁珠,并使用1 mL结合缓冲液清洗。 2.吸取适量人IgG,置于一个单独的管子中。 3.将人IgG与I-125标记的人IgG(30,000–100,000 cpm)混合。 4.使用结合缓冲液将人IgG与I-125标记的人IgG混合物稀释至100mL。 5.室温下孵育30分钟,使用闪烁计数器检测cpm值。 6.清洗磁珠(和包被层)4次,再次检测cpm值。 使用下述方程计算结合率%:(清洗后cpm值/清洗前cpm值)x100%。