The product Technical Bulletin is available is the ‘Documentation’ section above, under the ‘More Documents’ tab. A link has been provided below. These beads may be regenerated and reused multiple times. Due to varying sample characteristics, a finite number has not been established. However, more than 20 purifications are typical. It is recommended that the column be regenerated immediately after use by washing with at least 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. For column storage, wash the column with ten column volumes of 50% glycerol with TBS or PBS buffer containing 0.02% sodium azide. Then add another 5 mL of buffered glycerol containing 0.02% sodium azide and store at 2–8 °C or –20 °C without draining. Please see the link below to review additional protocols and product details:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/187/075/m8823bul-ms.pdf
M8823
Billes magnétiques Anti-FLAG® M2
affinity isolated antibody
Synonyme(s) :
Anticorps monoclonal ANTI-FLAG® M2 antibody produced in mouse, Résine FLAG® pour analyse à haut débit, Résine d’affinité Flag®, Résine d’affinité magnétique FLAG®, Anti-ddddk, Anti-dykddddk
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A propos de cet article
conjugate
magnetic beads
Quality Segment
antibody form
affinity isolated antibody
antibody product type
primary antibodies
clone
M2, monoclonal
form
suspension
shelf life
2 yr at -20 °C
analyte chemical class(es)
proteins
technique(s)
affinity chromatography: suitable, immunoprecipitation (IP): suitable, FLAG purification: suitable
bead size
20-75 μm
matrix
superparamagnetic iron impregnated 4% agarose bead, with an average diameter of 50 μm.
isotype
IgG1
capacity
≥0.6 mg/mL binding capacity
shipped in
wet ice
storage temp.
−20°C
General description
Application
Élution - peptide FLAG®, glycine, pH 3,5, peptide 3x FLAG®.
Pour en savoir plus sur nos produits, rendez-vous sur notre portail des applications des produits FLAG®.
Features and Benefits
- Réaliser des séparations très rapides
- Accélérer de manière significative les manipulations, par exemple les lavages répétitifs
- Traiter de nombreux échantillons à l'aide de plaques
Ce qui vous permet d'obtenir :
- Des expérimentations plus rapides
- Une meilleure reproductibilité
- Une quantification plus précise des protéines étudiées
Physical form
Legal Information
Disclaimer
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| description affinity isolated antibody | description 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) | description purified immunoglobulin, buffered aqueous glycerol solution | description 1.0-1.2 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4) |
| conjugate magnetic beads | conjugate unconjugated | conjugate agarose conjugate | conjugate unconjugated |
| clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal |
| antibody form affinity isolated antibody | antibody form purified immunoglobulin (Purified IgG1 subclass) | antibody form purified immunoglobulin | antibody form affinity isolated antibody |
| form suspension | form buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide) | form buffered aqueous glycerol solution | form buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4) |
| shipped in wet ice | shipped in dry ice | shipped in wet ice | shipped in wet ice |
| technique(s) immunoprecipitation (IP): suitable, affinity chromatography: suitable, FLAG purification: suitable | technique(s) western blot: 10 μg/mL (Protein A), FLAG purification: suitable | technique(s) FLAG purification: suitable, affinity chromatography: suitable, immunoprecipitation (IP): suitable | technique(s) FLAG purification: suitable |
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Classe de stockage
12 - Non Combustible Liquids
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
EPI (équipement de protection individuelle)
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
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where can I get the manual for these beads? is it possible to reuse them? if so, what is the protocol for the regeneration?
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Are these magnetic beads porous or no nonporous?
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These beads are porous.
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Bonjour Je souhaiterais que vous me confirmiez que les billes Billes magnétiques Anti-FLAG® M2 sont bien directement couplées à l'Anticorps anti flag ? Pouvez vous également me fournir un protocole d'immunocapture utilisant ces billes s'il vous plait ?
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The M2 Flag antibody is linked to the resin through a covalent linkage to one heavy chain. However, during the conjugation process, some Flag antibodies may also become bound to the resin through noncovalent attachments. To mitigate this, it is recommended to wash the resin using 0.1M Glycine followed by equilibration before use. This washing step ensures that the noncovalently linked antibodies are washed off. The protocol for this product can be found on the product page under more documents, where the Product Information sheet is available.
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Which antibody is used to bind to the beads in Anti-FLAG® M2 Magnetic Beads(M8823) and EZview™ Red ANTI-FLAG® M2 Affinity Gel(F2426)? I have ordered the antibody(F1804 Monoclonal ANTI-FLAG® M2 antibody produced in mouse). Is it this one?
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The antibody used for these items is proprietary information. ANTI-FLAG M2 Magnetic Beads consist of a mouse-derived, ANTI-FLAG M2 monoclonal antibody attached to superparamagnetic iron-impregnated 4% agarose beads. EZview Red ANTI-FLAG M2 Affinity Gel contains ANTI-FLAG M2 monoclonal antibody covalently attached to cross-linked 4% agarose beads. The ANTI-FLAG M2 antibody recognizes the FLAG octapeptide sequence (N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C) at the N-terminus, Met-N-terminus, or C-terminus locations of a fusion protein.
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Hello, I want to ask how many Flag-antibody on those beads?
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The specific number of antibodies per bead is not determined. The minimum amount of antibody in this product is 2.5 mg/mL of suspension. A range of 1.7 - 2.1 mL of suspension will yield 1 mL of packed resin. Both of these values are lot specific and reported in the Certificate of Analysis. Please see the link below to review a sample or lot specific Certificate:
https://www.sigmaaldrich.com/product/sigma/m8823#product-documentationHelpful?
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What is the maximum number of times that the ANTI-FLAG® M2 Magnetic Beads (M8823) can be reused?
What is the maximum number of times that the ANTI-FLAG® M2 Magnetic Beads (M8823) can be reused?
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The maximum number of times that the ANTI-FLAG® M2 Magnetic Beads (M8823) can be reused varies depending on the specific sample applied. It can range from one to twenty uses. Factors such as the cleanliness of the sample and the presence of substances that may clog the resin or cause nonspecific binding can affect the number of reuses. In typical scenarios, the beads are commonly reused for 3-5 times. It is essential to refer to the technical bulletin for specific situations where the beads cannot be reused, such as when using reagents containing SDS: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/187/075/m8823bul-ms.pdf
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Is there a protocol available for using the Anti-Flag M2 Magnetic Beads in a ChIP-Seq assay? Additionally, if the lysis buffer contains SDS and sodium deoxycholate at a concentration of 0.1%, will the presence of both denature or interfere with binding in the research process?
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The use of Anti-Flag M2 Magnetic Beads in a ChIP-Seq assay has not been validated. However, according to a citation provided in Stem Cells (PMID: 31348575), it suggests that these beads are compatible for such applications. Please note that M8823 is not recommended for use with Sodium Dodecyl Sulfate (SDS) or deoxycholate, as these components can denature the immobilized antibody and disrupt its binding to FLAG fusion proteins.
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Is Product No. CL4B200, Sepharose® CL-4B a suitable alternative for a negative bead control if magnetic beads without FLAG are not available? Are there any other recommendations for this purpose?
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For a negative control bead, any underivatized bead can essentially be used. While the Anti-FLAG® M2 Magnetic Beads (M8823) will work as a negative bead, it may not be suitable for magnetic applications. If a magnetic format is required, it is recommended to use polystyrene-based magnetized beads such as Product No. 49664, Micro particles based on polystyrene, magnetic. Alternatively, another option is to create a negative control using a non-FLAG tagged protein. This control can demonstrate the specificity of M8823 by showing its significant detection of FLAG-tagged proteins.
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Does M8823 have the capability to detect internal FLAG tags in addition to N and C terminal sequences?
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This paper (Zordan RE, Beliveau BJ, Trow JA, Craig NL, Cormack BP. Avoiding the ends: internal epitope tagging of proteins using transposon Tn7. Genetics. 2015 May;200(1):47-58. doi: 10.1534/genetics.114.169482. Epub 2015 Mar 5. PMID: 25745023; PMCID: PMC4423380) indicates that internal FLAG epiptopes were inserted into ORFs in yeast and detected by our F1804 antibody via Western blot. The biggest issue to overcome is the availability of the epitope for the antibody. If it is buried where it is not accessible in its native form, may not be possible. Denaturing the protein at least in this situation shows that the antibody can detect the protein.
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Are the M2 antibodies on the Anti-FLAG® M2 Magnetic Beads covalently bound?
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The M2 Flag antibody is linked to the resin through a covalent linkage to one heavy chain. However, during the conjugation process, some Flag antibodies may also become bound to the resin through noncovalent attachments. To mitigate this, it is recommended to wash the resin using 0.1M Glycine followed by equilibration before using it. This washing step ensures that the noncovalently linked antibodies are washed off.
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