Dynabeads™ His-Tag Isolation and Pulldown
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Dynabeads™ His-Tag Isolation and Pulldown
Invitrogen™

Dynabeads™ His-Tag Isolation and Pulldown

Dynabeads™ His-Tag Isolation and Pulldown beads are 1-μm monosized, monodispersed, superparamagnetic beads with an NTA-Cobalt-based ligand which has a high了解更多信息
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货号数量
10104D10 mL
10103D2 mL
货号 10104D
价格(CNY)
9,313.00
飞享价
Ends: 31-Dec-2025
12,417.00
共减 3,104.00 (25%)
Each
添加至购物车
数量:
10 mL
价格(CNY)
9,313.00
飞享价
Ends: 31-Dec-2025
12,417.00
共减 3,104.00 (25%)
Each
添加至购物车

Dynabeads™ His-Tag Isolation and Pulldown beads are 1-μm monosized, monodispersed, superparamagnetic beads with an NTA-Cobalt-based ligand which has a high affinity towards His-tags and similar structures on molecular targets. Their small but uniform bead size presents a high surface area per mg beads and a correspondingly high capacity for the target molecule. They enable single-step isolation of tagged molecules in manual or automated workflows that helps ensure good recovery of the target. Dynabeads His-Tag Isolation and Pulldown beads can also be a great starting point for other advanced experimental approaches where solid magnetic support is needed.

Benefits of Dynabeads His-Tag and Pulldown beads include:
Performance—high sensitivity and reproducibility securing consistent results
Efficiency—fast binding kinetics and high binding capacity
Flexibility—flexible protocols with gentle and efficient liquid-phase reaction kinetics
Automation-ready—automation protocols available with the KingFisher Purification instruments

Good signal-to-noise for numerous applications
Dynabeads His-Tag Isolation and Pulldown beads are an ideal tool for isolation of tagged recombinant proteins. They are also suited for other applications with high signal-to-noise requirements. Using the principles of immobilized metal affinity binding together with the inert surface of Dynabeads magnetic beads helps ensure a clean capture with extremely low carryover of unwanted proteins. Since the bead surface is loaded with cobalt ions, it is also possible to make a stable and lasting bond with the target molecule through oxidation if the downstream applications require it. The beads are added to the solution containing the target and mixed for 5-10 minutes. After washing, the target is eluted off and ready for downstream applications.

More than just a pulldown
In addition to being a great tool for protein pulldown and isolation, the Dynabeads His-Tag Isolation and Pulldown beads can serve as a platform for target immobilization. The bead carries cobalt Co2+ ions, so it is possible to cement the bond between the bead and the target just by using mildly oxidative conditions, such as 10 mM hydrogen peroxide for 30–60 minutes. This oxidizes cobalt to the Co3+ state, making the connection between the bead and the target permanent. It will not elute in the commonly used elution buffers for IMAC. This can later be used for applications requiring a strongly bound target, without the risk of reduced target yield.

Automation-ready for use with KingFisher purification systems
The 1-μm magnetic beads are ideal for high-throughput enrichment. This process can be automated using any of our KingFisher sample purification systems or liquid handlers.

Commercial supply 
Our manufacturing sites are ISO 13485-certified. Bead characteristics and manufacturing conditions help ensure consistency and batch reproducibility making them an ideal choice for commercial supply. If you are in the process of customizing Dynabeads His-Tag Isolation and Pulldown beads in a commercial product or service or need a larger bulk volume of this product, please contact us at oemdynal@lifetech.com or see our Dynabeads OEM page.

Learn more about Dynabeads magnetic beads and KingFisher Sample Purification Systems:
See our immunoprecipitation selection table, data, and references
Dynabeads magnetic beads selection guide
See magnets for Dynabeads magnetic beads separations
KingFisher automation protocols
Watch a video about the KingFisher Flex instrument

仅供科研使用。不可用于诊断程序。
规格
结合属性1.6 mg of a 28 kDa histidine-tagged protein/mL (40 mg) beads
认证/合规ISO9001 和 ISO13485
颜色Brown
最大浓度40 mg/mL
直径(公制)1 μm
最终产品类型His-Tagged Protein
适用于(应用)Isolation of His-tagged Recombinant Proteins
适用于(设备)KingFisher Sample Purification System, DynaMag magnets
产品规格Magnetic beads
高通量能力兼容高通量应用
配基类型Nitrilotriacetic acid (NTA)
材料磁性聚苯乙烯
数量10 mL
调控状态For Research Use Only
样品类型Cell Cultures, Tissue Digests, Intact Proteins, Cell Lysates, Cell Extracts
有效期自生产之日起 36 个月
运输条件室温
足够用于200 Tests
表面功能基团Cobalt-based immobilized metal affinity
靶标His-tagged proteins
产品线Dynabeads™
类型磁珠
Unit SizeEach
内容与储存
Dynabeads His-Tag Isolation and Pulldown beads are supplied in 20% ethanol. Store at 2°C to 8°C.

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%。