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M8823

Anti-FLAG® M2 Magnetic Beads

affinity isolated antibody

Anti-FLAG® M2 Magnetic Beads
1 of 1 reviewers received a sample product or took part in a promotion

Synonim(y):

Monoclonal ANTI-FLAG® M2, FLAG® magnetic affinity resin, FLAG® resin for high throughput, Flag® Affinity resin, Anti-ddddk, Anti-dykddddk

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Do Państwa/SKUDostępnośćCena netto
1 mL
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3230,00 zł
5 mL
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9660,00 zł
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24 570,00 zł

Informacje o tej pozycji

NACRES:
NA.32
UNSPSC Code:
12352203
Conjugate:
magnetic beads
Clone:
M2, monoclonal
Application:
IP, affinity chromatography
Citations:
1017

3230,00 zł


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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

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

Anti-FLAG M2 Magnetic Beads are 4% agarose beads bound with the Anti-FLAG M2 (mouse monoclonal) antibody. The M2 antibody recognizes the FLAG sequence at the N-terminus, Met-N-terminus and C-terminus. This alows for detection and capture of fusion proteins containing a FLAG peptide sequence.

Application

Suitable for immunoprecipitation procedures.

Elution - FLAG® peptide, Glycine, pH 3.5, 3x FLAG® peptide

Learn more product details in our FLAG® application portal.

Features and Benefits

The magnetic properties allow for:
- Very rapid separation
- Significantly accelerated manipulations, such as repetitive washings
- Processing of multiple samples performed in plate formats
This leads to:
- Faster experimentation
- Better reproducibility
- More accurate quantitation of the proteins of interest

Physical form

Supplied as a 50% suspension in 50% glycerol with 10mM sodium phosphate, 150mM sodium chloride, pH 7.4 and 0.02% (w/v) sodium azide (PBA/A).

Legal Information

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

Disclaimer

FLAG® Affinity Gels, FLAG® tag, 3x FLAG® tag, DYKDDDDK tag
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F1804F2426F4049
description

affinity isolated antibody

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)

description

clone M2

description

clone M2, purified immunoglobulin, buffered aqueous solution

antibody form

affinity isolated antibody

antibody form

affinity isolated antibody

antibody form

-

antibody form

purified immunoglobulin

clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

conjugate

magnetic beads

conjugate

unconjugated

conjugate

-

conjugate

FITC conjugate

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)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

technique(s)

direct immunofluorescence: 10 μg/mL using mammalian cells fixed with methanol:acetone

shipped in

wet ice

shipped in

wet ice

shipped in

wet ice

shipped in

dry ice


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Klasa składowania

12 - Non Combustible Liquids

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)



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Certyfikaty analizy (CoA)

Lot/Batch Number

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Questions

1–10 of 13 Questions  
  1. where can I get the manual for these beads? is it possible to reuse them? if so, what is the protocol for the regeneration?

    1 answer
    1. 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

      Helpful?

  2. Are these magnetic beads porous or no nonporous?

    1 answer
    1. These beads are porous.

      Helpful?

  3. 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 ?

    1 answer
    1. 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.

      Helpful?

  4. 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?

    1 answer
    1. 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.

      Helpful?

  5. Hello, I want to ask how many Flag-antibody on those beads?

    1 answer
    1. 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-documentation

      Helpful?

  6. 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?

    1 answer
    1. 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

      Helpful?

  7. 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?

    1 answer
    1. 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.

      Helpful?

  8. 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?

    1 answer
    1. 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.

      Helpful?

  9. Does M8823 have the capability to detect internal FLAG tags in addition to N and C terminal sequences?

    1 answer
    1. 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.

      Helpful?

  10. Are the M2 antibodies on the Anti-FLAG® M2 Magnetic Beads covalently bound?

    1 answer
    1. 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.

      Helpful?

1–10 of 13 Questions  

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