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Merck
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安全信息

A2220

Millipore

抗-FLAG® M2亲和凝胶

purified immunoglobulin, buffered aqueous glycerol solution

别名:

单克隆抗-FLAG® M2 小鼠抗, 抗-FLAG® M2亲和琼脂糖凝胶, 抗 ddddk, 抗 dykddddk

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About This Item

UNSPSC代码:
12352203
NACRES:
NA.32

偶联物

agarose conjugate

质量水平

200

抗体形式

purified immunoglobulin

抗体产品类型

primary antibodies

克隆

M2, monoclonal

形式

buffered aqueous glycerol solution

分析物化学类别

proteins

技术

affinity chromatography: suitable
immunoprecipitation (IP): suitable

基质

(4% agarose bead; 45-165μm bead size)

同位素/亚型

IgG1

容量

>0.6 mg/mL, resin binding capacity (FLAG-BAP)

运输

wet ice

储存温度

−20°C

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相关类别

一般描述

抗-FLAG M2亲和凝胶是一种可与琼脂糖共价连接的小鼠单克隆抗体。该抗体可在融合蛋白的N末端、Met-N末端、C末端和内部位置与FLAG结合。结合是不依赖于钙的。

洗脱产物 - FLAG®肽，甘氨酸，pH3.5，3x FLAG®肽
FLAG® peptide, Glycine, pH3.5, 3x FLAG® peptide

免疫原

DYKDDDDK

应用

抗-FLAG®M2亲和凝胶已用于western blotting、免疫沉淀以及FLAG融合蛋白的纯化。

请访问我们的FLAG®应用门户网站，了解更多产品详情。

外形

悬浮于含有叠氮化物作为防腐剂以及50％甘油的缓冲盐溶液中

其他说明

2024年CiteAb奖——成功的帕金森研究供应商(2024 CiteAb Award Winner for Supplier Succeeding in Parkinson′s Research)

法律信息

ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany

免责声明

FLAG™ tag, 3x FLAG™, DYKDDDDK tag

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WGK

WGK 1

闪点(°F)

Not applicable

闪点(°C)

Not applicable

法规信息

常规特殊物品

分析证书(COA)

输入产品批号来搜索 分析证书(COA) 。批号可以在产品标签上"批“ （Lot或Batch）字后找到。

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  1. Is my lysis buffer compatible with Product No. A2220, ANTI-FLAG® M2 Affinity Gel?

    The A2220 product information sheet (under Documents, above) contains a reagent compatibility table located on page 6. We do not recommend addition of SDS or reducing agents such as DTT, DTE or 2-mercaptoethanol to the resin. If you have a detergent listed in the table in a higher than recommended concentration, we recommend trying to dilute the sample before applying to the resin.

  2. How should I elute my protein when using Product No. A2220, ANTI-FLAG® M2 Affinity Gel?

    Elution with the peptide is the most gentle method. Acid elution (0.1 M glycine-HCL pH 3.5) is a more stringent method of elution, and should be evaluated for its effect on your protein if it is to be used in downstream applications. Boiling the resin in sample buffer is the most denaturing condition. If this condition is used, the resin cannot be re-used, due to the presence of SDS and/or reducing agents. The elution information can be viewed on A2220 product information sheet (under Documents, above).

  3. When using Product No. A2220, ANTI-FLAG® M2 Affinity Gel, I see bands at 20-25 kDa and 50-60 kDa appearing in my Westerns that are not my FLAG®-tagged protein. How can I prevent this?

    As a result of the conjugation, there may be some M2 antibody that is not conjugated to the resin, but is associated with the resin and may appear in acid elutions as heavy and light chain when using the anti-mouse IgG conjugated secondary antibody. We recommend an acid wash (0.1 M glycine-HCL pH 3.5) and neutralization of the resin (do not allow the acid wash to sit on the resin longer than 20 minutes) prior to applying the lysate.  Another way to avoid this is to use a directly conjugated FLAG® antibody for detection such as product A8592 ANTI-FLAG® M2 HRP, or the rabbit anti-FLAG® polyclonal antibody, F7425.

  4. I am using Product No. A2220, ANTI-FLAG® M2 Affinity Gel, and have a lot of non-specific proteins that are eluting with my FLAG®-tagged protein.  How can I get rid of these?

    The product bulletin for Product A2220, ANTI-FLAG® M2 affinity gel indicates:Pre-clear lysate with Mouse IgG-Agarose (Product A0919) to remove nonspecific binding proteins. Alternatively, you can use the unconjugated resin (Product 4B200) for this purpose. Other methods to remove non-specific binding from the resin would be to increase the stringency of the washes by increasing salt concentration (the resin can tolerate up to 1M NaCl) or including detergents that are compatible with the resin.

  5. When using ANTI-FLAG M2 Affinity Gel, Product A2220, should I use a 3X FLAG peptide or a 1X FLAG peptide to elute my protein?

    If you have a 3X FLAG-tagged protein, then you will need to use the 3X FLAG peptide.  If you have a 1X FLAG-tagged protein, you can use the 1X FLAG peptide or the 3X FLAG peptide.  We have not noticed a significant  difference in elution efficiency by using a 3X FLAG peptide on a 1X FLAG-tagged protein. 

  6. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  9. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Yu Ti Cheng et al.
Proceedings of the National Academy of Sciences of the United States of America, 108(35), 14694-14699 (2011-08-30)
The nucleotide-binding domain and leucine-rich repeats containing proteins (NLRs) serve as immune receptors in both plants and animals. Overaccumulation of NLRs often leads to autoimmune responses, suggesting that the levels of these immune receptors must be tightly controlled. However, the
Nora Nonne et al.
Nucleic acids research, 38(4), e20-e20 (2009-12-04)
MicroRNAs (miRNAs) bind to Argonaute proteins, and together they form the RISC complex and regulate target mRNA translation and/or stability. Identification of mRNA targets is key to deciphering the physiological functions and mode of action of miRNAs. In mammals, miRNAs
Michelle F Green et al.
The Journal of biological chemistry, 286(32), 28066-28079 (2011-06-15)
Ca(2+)/calmodulin-dependent protein kinase kinase β (CaMKKβ) is a serine/threonine-directed kinase that is activated following increases in intracellular Ca(2+). CaMKKβ activates Ca(2+)/calmodulin-dependent protein kinase I, Ca(2+)/calmodulin-dependent protein kinase IV, and the AMP-dependent protein kinase in a number of physiological pathways, including
Xinna Zhang et al.
The EMBO journal, 30(11), 2177-2189 (2011-04-28)
Tumour suppressor p53 levels in the cell are tightly regulated by controlled degradation through ubiquitin ligases including Mdm2, COP1, Pirh2, and ARF-BP1. The ubiquitination process is reversible via deubiquitinating enzymes, such as ubiquitin-specific peptidases (USPs). In this study, we identified
William G Roach et al.
The Biochemical journal, 403(2), 353-358 (2007-02-06)
Insulin stimulation of the trafficking of the glucose transporter GLUT4 to the plasma membrane is controlled in part by the phosphorylation of the Rab GAP (GTPase-activating protein) AS160 (also known as Tbc1d4). Considerable evidence indicates that the phosphorylation of this
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商品

ANTI-FLAG® M2 Magnetic Beads

The FLAG® Expression System is a proven method to express, purify and detect recombinant fusion proteins. Sigma®, the proven provider of FLAG®, now offers a magnetic bead for immunoprecipitation, protein purification, and the study of protein-protein interactions. The ANTI-FLAG® M2 Magnetic Bead is composed of murine derived, anti-FLAG® M2 monoclonal antibody attached to superparamagnetic iron impregated 4% agarose beads, with an average diameter of 50 µm. The M2 antibody is capable of binding to fusion proteins containing a FLAG peptide sequence at the N-terminus, Met-N-terminus, or C-terminus locations in mammalian, bacterial, and plant extracts.

ANTI-FLAG® M2磁珠

FLAG®表达系统是一种表达、纯化和检测重组融合蛋白的成熟方法。作为FLAG®的可靠供应商，Sigma®现提供FLAG标签抗体标记的免疫磁珠，用于免疫沉淀、蛋白纯化和蛋白互作研究。ANTI-FLAG® M2磁珠由ANTI-FLAG® M2鼠单克隆抗体与超顺磁铁包覆的4%琼脂糖微珠偶联而成，平均粒径50 µM。M2抗体可结合哺乳动物、细菌和植物提取物融合蛋白N端、Met-N端或C端的FLAG®肽。

实验方案

Immunoprecipitation of FLAG Fusion Proteins Using Monoclonal Antibody Affinity Gels

Protocol for immunoprecipitation (IP) of FLAG fusion proteins using M2 monoclonal antibody 4% agarose affinity gels

采用单克隆抗体亲和凝胶进行的FLAG融合蛋白免疫沉淀_蛋白纯化-默克生命科学

免疫沉淀（IP）可用于高效、高产量分离和纯化与FLAG®肽标签融合的蛋白质。此过程采用ANTI-FLAG® M2亲和凝胶进行，后者是一种与琼脂糖树脂共价结合的高度特异性单克隆抗体。

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Protein Purification

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