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Merck

A2220

抗FLAG® M2抗体 アフィニティーゲル

purified immunoglobulin, buffered aqueous glycerol solution

ANTI-FLAG® M2 Affinity Gel

別名:

モノクローナル抗FLAG® M2抗体 マウス宿主抗体, 抗FLAG®M2抗体アフィニティアガロースゲル, 抗ddddk, 抗dykddddk

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1 ML

￥61,360

5 ML

￥182,400

10 ML

￥324,750

25 ML

￥730,400

￥61,360

単価￥76,700割引20%

本日発送予定詳細

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この商品について

NACRES:
NA.32
UNSPSC Code:
12352203

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製品名

抗FLAG® M2抗体 アフィニティーゲル, purified immunoglobulin, buffered aqueous glycerol solution

conjugate

agarose conjugate

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

M2, monoclonal

form

buffered aqueous glycerol solution

analyte chemical class(es)

proteins

technique(s)

affinity chromatography: suitable
immunoprecipitation (IP): suitable

matrix

(4% agarose bead; 45-165μm bead size)

isotype

IgG1

capacity

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

shipped in

wet ice

storage temp.

−20°C

Quality Level

200

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当該品目
F1804A4596F3165
抗FLAG® M2抗体 アフィニティーゲル purified immunoglobulin, buffered aqueous glycerol solution

Millipore

A2220

抗FLAG® M2抗体 アフィニティーゲル

モノクローナル抗FLAG® M2抗体 マウス宿主抗体 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)

Sigma-Aldrich

F1804

モノクローナル抗FLAG® M2抗体 マウス宿主抗体

抗FLAG® M1抗体-アガロース結合アフィニティーゲル

Millipore

A4596

抗FLAG® M1抗体-アガロース結合アフィニティーゲル

モノクローナル抗FLAG® M2抗体 マウス宿主抗体 clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)

Sigma-Aldrich

F3165

モノクローナル抗FLAG® M2抗体 マウス宿主抗体

antibody form

purified immunoglobulin

antibody form

affinity isolated antibody

antibody form

-

antibody form

purified immunoglobulin (Purified IgG1 subclass)

conjugate

agarose conjugate

conjugate

unconjugated

conjugate

agarose conjugate

conjugate

unconjugated

clone

M2, monoclonal

clone

M2, monoclonal

clone

-

clone

M2, monoclonal

shipped in

wet ice

shipped in

wet ice

shipped in

-

shipped in

dry ice

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

technique(s)

-

technique(s)

-

technique(s)

western blot: 10 μg/mL (Protein A)

Application

抗-FLAG® M2アフィニティゲルは、ウェスタンブロッティング、免疫沈降、およびFLAG融合タンパク質の精製に使用されています。[1][2][3]

製品の詳細につきましては、FLAG®アプリケーションポータルをご覧ください。

Disclaimer

FLAG® タグ、3x FLAG®、DYKDDDDK タグ

General description

FLAG® ペプチド、グリシン、pH3.5、3 x FLAG® ペプチド
抗-FLAG M2アフィニティゲルは、マウスモノクローナル抗体を結合させたアガロースです。本抗体は、融合タンパク質のN-末端、Met-N-末端、C-末端、および内部に位置するFLAGに結合します。結合はカルシウム非依存性です。

溶出 - FLAG®ペプチド、グリシン、pH 3.5、3x FLAG®ペプチド

Immunogen

DYKDDDDK

Other Notes

パーキンソン病の研究に貢献したサプライヤーに贈られる2024 CiteAb賞の受賞者＠′＠

Physical form

保存料としてアジド、および 50％グリセロールを含む緩衝食塩水中に懸濁

Legal Information

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

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Related product

製品番号
詳細
価格

保管分類

10 - Combustible liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

最新バージョンのいずれかを選択してください：

試験成績書（COA）

Lot/Batch Number
0000508520
0000508518
0000496338
0000496338
0000491728

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特定のバージョンが必要な場合は、ロット番号またはバッチ番号で特定の証明書を検索できます。

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以前この製品を購入いただいたことがある場合

文書ライブラリで、最近購入した製品の文書を検索できます。

文書ライブラリにアクセスする

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
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
Joshua M Baughman et al.
Nature, 476(7360), 341-345 (2011-06-21)
Mitochondria from diverse organisms are capable of transporting large amounts of Ca(2+) via a ruthenium-red-sensitive, membrane-potential-dependent mechanism called the uniporter. Although the uniporter's biophysical properties have been studied extensively, its molecular composition remains elusive. We recently used comparative proteomics to
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
Qian Yang et al.
Journal of virology, 93(16) (2019-06-07)
Hemoglobin is an important oxygen-carrying protein and plays crucial roles in establishing host resistance against pathogens and in regulating innate immune responses. The hemoglobin subunit beta (HB) is an essential component of hemoglobin, and we have previously demonstrated that the
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資料

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.

プロトコル

モノクローナル抗体アフィニティーゲルを使用したFLAG®融合タンパク質の免疫沈降

M2モノクローナル抗体4%アガロースアフィニティーゲルを使用したFLAG®融合タンパク質の免疫沈降（IP）のためのプロトコル

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

関連コンテンツ

Protein Research Guide

Find protein research tools to prepare, isolate, and analyze proteins. Organized by how to extract, protect, purify, enrich, modify, and quantify proteins.

Protein Purification

Protein purification techniques, reagents, and protocols for purifying recombinant proteins using methods including, ion-exchange, size-exclusion, and protein affinity chromatography.

タンパク質精製

イオン交換、サイズ排除、タンパク質アフィニティークロマトグラフィーなどの方法を用いた組換えタンパク質精製のためのタンパク質精製技術、試薬、プロトコル。

EZview Red Protein A and ANTI-FLAG M2 Affinity Gels

EZviewTM Red Protein A and ANTI-FLAG® M2 Affinity Gels: Immunoprecipitation with Enhanced Visibility Affinity Beads - Technical Article - July 2001

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質問

1–10/11 質問  

  1. How do I equilibrate the resin and how can I clean it for re-use?

    1 回答

    1. Detailed equilibration and regeneration protocols for this product are provided in the Product Information Sheet:
      http://sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/288/183/a2220bul-ms.pdf

      役に立ちましたか？

  2. May I ask a question about the product? Does "1mL" mean 1 mL Slurry plus 50% glycerol, which is 2 mL in total? Same as other (e.g., 10 mL = 50% slurry with total 20 mL volume)

    1 回答

    1. The "1mL" indicates that this product is supplied at a volume of 1 milliliter. This ANTI-FLAG M2 affinity gel is supplied as a 50% suspension in 50% glycerol with 10 mM sodium phosphate and 150 mM sodium chloride, pH 7.4, containing 0.02% (w/v) sodium azide (PBS/A).

      役に立ちましたか？

  3. What is the bead size for this product?

    1 回答

    1. As stated in the 'PROPERTIES' section of the product page, the bead size range is 45-165 μm.

      役に立ちましたか？

  4. Is there a generic agarose bead available for purchase that can be used as a control to test for non-specific binding, in the case that something is binding to the agarose bead itself rather than the tag? If it is not listed on the site, is there a possibility of obtaining generic agarose beads through alternative means? Alternatively, could any information be provided to determine if another provider's generic agarose beads are produced in a similar manner for this specific purpose?

    1 回答

    1. Yes, there are two options available for a generic agarose bead as a control to test for non-specific binding. One option is Product Number. 4B200, Sepharose® 4B, which can be used for sample preclearing and is equivalent to the beads in the FLAG affinity product. The other option is Product Number: A0919, Mouse IgG−Agarose, which can serve as a conjugated antibody bead control.

      役に立ちましたか？

  5. Is there a protocol for regeneration?

    1 回答

    1. Please see the link below to the product data-sheet. Instructions state that the column should be regenerated immediately after use by washing with three column volumes of 0.1 M glycine HCI, pH 3.5. The column should be immediately re-equilibrated in TBS until the effluent is at neutral pH:

      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/288/183/a2220bul-ms.pdf

      役に立ちましたか？

  6. 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?

    1 回答

    1. 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.

      役に立ちましたか？

  7. 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?

    1 回答

    1. 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.

      役に立ちましたか？

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

    1 回答

    1. 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.

      役に立ちましたか？

  9. 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?

    1 回答

    1. 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.

      役に立ちましたか？

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

    1 回答

    1. 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).

      役に立ちましたか？

1–10/11 質問  
2件のレビュー このアクションによりレビューへと移動します。

レビュー

アクティブなフィルタ

  1. Baton Rouge
    • 1件のレビュー
    • 1件の投票
    星5／5個です。

    Works well

    This resin works well at purifying FLAG-tagged proteins very specifically if you do the correct amount of washes (5).

    Googleで翻訳する

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  2. 1件の評価のみのレビュー

ライフサイエンス、有機合成、材料科学、クロマトグラフィー、分析など、あらゆる分野の研究に経験のあるメンバーがおります。.

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