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F3165

ANTI-FLAG® M2 antibody, Mouse monoclonal

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)

ANTI-FLAG® M2 antibody, Mouse monoclonal

Synonym(s):

Anti-ddddk, Anti-dykddddk, Monoclonal ANTI-FLAG® M2

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About This Item

NACRES:
NA.32
UNSPSC Code:
12352203
Conjugate:
unconjugated
Clone:
M2, monoclonal
Application:
WB
Citations:
8404
Pricing and availability is not currently available.
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biological source

mouse

Quality Level

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

General description

Anti-Flag M2 antibody is used for the detection of Flag fusion proteins. This monoclonal antibody 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 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.

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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This Item
F1804A9469A2220
conjugate

unconjugated

conjugate

unconjugated

conjugate

alkaline phosphatase conjugate

conjugate

agarose conjugate

biological source

mouse

biological source

mouse

biological source

mouse

biological source

-

antibody form

purified immunoglobulin (Purified IgG1 subclass)

antibody form

affinity isolated antibody

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

species reactivity

all

species reactivity

all

species reactivity

all

species reactivity

-

clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

technique(s)

western blot: 10 μg/mL (Protein A)

technique(s)

-

technique(s)

indirect ELISA: 1:20,000

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable


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Storage Class

12 - Non Combustible Liquids

flash_point_f

Not applicable

flash_point_c

Not applicable



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Related Content

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


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
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
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



Global Trade Item Number

SKUGTIN
F3165-.2MG04061835518128
F3165-5MG04061833613542
F3165-1MG04061835512522

Questions

1–7 of 7 Questions  
  1. What is the recommended short-term storage condition for FLAG M2 antibody — is 4°C acceptable?

    1 answer
    1. To enhance stability & functional consistency, store the undiluted antibody at −20 °C in working aliquots. Repeated freezing and thawing is not recommended.

      Helpful?

  2. What is the recommended dilution for immunocytochemistry for this product ?

    1 answer
    1. This product has not been validated for immunocytochemistry applications.

      Helpful?

  3. 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?

  4. 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?

  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?

  6. 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?

  7. 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?

Reviews

Active Filters

  1. New Jersey
    • Review 1
    • Votes 15
    5 out of 5 stars.

    Great Anti Flag primary

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

    Helpful?