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F3165

Sigma-Aldrich

Monoclonal ANTI-FLAG® M2

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)

Monoclonal ANTI-FLAG® M2

Synonym(s):

Anti-ddddk, Anti-dykddddk

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$607.00
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$1,220.00
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$3,540.00

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.32

Key Documents

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biological source

mouse

Quality Level

200

conjugate

unconjugated

antibody form

purified immunoglobulin (Purified IgG1 subclass)

antibody product type

primary antibodies

clone

M2, monoclonal

form

buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)

purified by

using Protein A

species reactivity

all

concentration

3.8-4.2 mg/mL

technique(s)

western blot: 10 μg/mL (Protein A)

isotype

IgG1

immunogen sequence

DYKDDDDK

shipped in

dry ice

storage temp.

−20°C

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This Item
F9291A9594A8592
Monoclonal ANTI-FLAG® M2 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

Monoclonal ANTI-FLAG® M2

Monoclonal ANTI-FLAG® BioM2 antibody produced in mouse clone M2, purified immunoglobulin, buffered aqueous glycerol solution

Sigma-Aldrich

F9291

Monoclonal ANTI-FLAG® BioM2 antibody produced in mouse

Monoclonal ANTI-FLAG® M2-Cy3™ antibody produced in mouse clone M2, purified immunoglobulin, buffered aqueous solution (Supplied as a solution in 10 mM sodium phosphate)

Sigma-Aldrich

A9594

Monoclonal ANTI-FLAG® M2-Cy3™ antibody produced in mouse

Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody produced in mouse clone M2, purified immunoglobulin, buffered aqueous glycerol solution

Sigma-Aldrich

A8592

Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody produced in mouse

clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

conjugate

unconjugated

conjugate

biotin conjugate

conjugate

CY3 conjugate

conjugate

peroxidase conjugate

biological source

mouse

biological source

mouse

biological source

mouse

biological source

mouse

antibody form

purified immunoglobulin (Purified IgG1 subclass)

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

species reactivity

all

species reactivity

all

species reactivity

all

species reactivity

all

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 (Supplied as a solution in 10 mM sodium phosphate)

form

buffered aqueous glycerol solution

General description

Anti-Flag M2 antibody is used for the detection of Flag fusion proteins. This monoclonal antibody is produced in mouse and recognizes the FLAG sequence at the N-terminus, Met N-terminus, and C-terminus. The antibody is also able to recognize FLAG at an internal site. M2, unlike M1 antibody is not Calcium dependent.
F3165 is affinity purified using Protein A resin, so it contains not only the anti-FLAG M2 antibody but also small amounts of native mouse IgG, increasing its sensitivity in most applications.
Method of purification - Protein A

Immunogen

FLAG; peptide sequence DYKDDDDK

Application

Monoclonal ANTI-FLAG® M2 antibody produced in mouse has been used in:
  • immunoblotting
  • immunoprecipitation
  • immunocytochemistry
  • immunofluorescence
  • ELISA
  • EIA
  • chromatin immunoprecipitation
  • electron microscopy
  • flow cytometry
  • supershift assays

Browse additional application references in our FLAG® Literature portal.

Preparation Note

Dilute the antibody solution from 0.5-10 ug/mL in Tris Buffered Saline, pH 8.0, with 3% nonfat milk
Store the undiluted antibody at –20 °C in working aliquots. Repeated freezing and thawing is not recommended.
Note: Overtime, small amounts of purified antibodies can precipitate from solution due to intermolecular hydrophobic interactions. If a precipitate is observed in this product, briefly centrifuge the vial to pellet the precipitate. Withdraw the desired volume of antibody solution from the clear supernatant for use. This should not alter the performance of the purified antibody in Western blot or immunoprecipitation applications.

Other Notes

2024 CiteAb Award Winner for Supplier Succeeding in Parkinson′s Research

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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Storage Class

12 - Non Combustible Liquids

wgk_germany

nwg

flash_point_f

Not applicable

flash_point_c

Not applicable

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Certificates of Analysis (COA)

Lot/Batch Number
0000493941
0000493938
0000493940
0000493940
0000481877

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Casey C Fowler et al.
Nature communications, 10(1), 3684-3684 (2019-08-17)
Bacterial toxins with an AB5 architecture consist of an active (A) subunit inserted into a ring-like platform comprised of five delivery (B) subunits. Salmonella Typhi, the cause of typhoid fever, produces an unusual A2B5 toxin known as typhoid toxin. Here
T P Molitor et al.
Oncogenesis, 2, e48-e48 (2013-06-05)
The vaccinia-related kinases (VRKs) comprise a branch of the casein kinase family. VRK1, a ser/thr kinase with a nuclear localization, is the most well-studied paralog and has been described as a proproliferative protein. In lower eukaryotes, a loss of VRK1
Hong Zhu et al.
Molecular biology of the cell, 24(11), 1619-1637 (2013-04-12)
Charcot-Marie-Tooth (CMT) disease is an inherited neurological disorder. Mutations in the small integral membrane protein of the lysosome/late endosome (SIMPLE) account for the rare autosomal-dominant demyelination in CMT1C patients. Understanding the molecular basis of CMT1C pathogenesis is impeded, in part
Fangzhi Tan et al.
Nature communications, 10(1), 3733-3733 (2019-08-21)
Hearing loss is the most common sensory disorder. While gene therapy has emerged as a promising treatment of inherited diseases like hearing loss, it is dependent on the identification of gene delivery vectors. Adeno-associated virus (AAV) vector-mediated gene therapy has
Wenjiao Li et al.
Cell death and differentiation, 26(8), 1379-1395 (2018-10-14)
RASSF1A (Ras association domain family 1 isoform A) is a tumor suppressor and frequently inactivated by promoter hypermethylation in hepatocellular carcinoma (HCC). Autophagy is to degrade misfolded or aggregated proteins and dysfunctional organelles. Autophagy defects enhance oxidative stress and genome
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Comparison of elution techniques for small-scale protein purification of FLAG® tag proteins using anti-FLAG® M2 magnetic beads.

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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

Data Sheet - F3165
1 Review This action will navigate to reviews.

Questions

1–5 of 5 Questions  

  1. Is there a recommended dilution for capture (sandwich) based ELISA?

    1 answer

    1. Monoclonal ANTI-FLAG® M2 may be used in EIA procedures. Typically, a fusion protein containing a FLAG® peptide sequence is directly adsorbed (or otherwise presented) within the wells of a multiwell polystyrene plate. The Monoclonal ANTI-FLAG® M2 antibody may be diluted up to 1:50,000 for subsequent incubation within the plate wells. Detection may be accomplished using Anti-Mouse IgG-Peroxidase (Cat. No. A9044) or equivalent, diluted 1:10,000, followed by an appropriate substrate for visualization.

      Dilutions for use as the capture antibody in an ELISA have not been evaluated. We would recommend that the end-user optimize the protocol following the recommendations found here:
      https://www.sigmaaldrich.com/technical-documents/protocol/protein-biology/elisa/elisa-procedures#antigen_coating

      Helpful?

  2. What is the recommended dilution for conducting western blotting using this antibody?

    1 answer

    1. The recommended dilution for Western Blot is 10 µg/mL. Please see the link below to review additional information, including protocols:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/274/912/f3165dat-ms.pdf

      Helpful?

  3. What type of light chain is present on the FLAG M2 antibody?

    1 answer

    1. Test has not been conducted to determine whether the light chains of the antibody F3165, Monoclonal ANTI-FLAG® M2 antibody produced in mouse, are kappa or lambda. However, based on customer feedback, it appears that the M2 monoclonal antibody has lambda light chains and not kappa. A probe with anti-kappa did not yield a signal, while anti-lambda produced a good signal.

      Helpful?

  4. What is the recommended dilution for conducting immunoprecipitation (IP) using the M2 antibody?

    1 answer

    1. The general starting recommendation for conducting immunoprecipitation (IP) using the F3165, anti-FLAG M2 antibody, is 10 μg/mL.

      Helpful?

  5. What is the difference between F1804 and F3165 product?

    1 answer

    1. Product F1804 and F3165 are both Monoclonal ANTI-FLAG M2 antibodies produced in mouse Clone M2, with the same starting material. The distinction between the products lies in their purification methods. F3165 is immunoglobulin purified, while F1804 is immunogen affinity purified. F1804, being more purified, is expected to exhibit reduced background and less non-specific staining compared to F3165. Both products are suitable for western blotting, immunofluorescence, and immunoprecipitation.

      Helpful?

Reviews

Active Filters

  1. New Jersey
    • Review 1
    • Votes 13
    5 out of 5 stars.

    Great Anti Flag primary

    Works well for our western blot analysis of flag tagged recombinant proteins

    Helpful?

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