To enhance stability & functional consistency, store the undiluted antibody at −20 °C in working aliquots. Repeated freezing and thawing is not recommended.
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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0.2 MG
$582.00
1 MG
$1,200.00
5 MG
$3,440.00
About This Item
NACRES:
NA.32
UNSPSC Code:
12352203
Conjugate:
unconjugated
Clone:
M2, monoclonal
Application:
WB
Citations:
8404
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biological source
mouse
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
Quality Level
Related Categories
1 of 4
This Item | F9291 | A9594 | A8592 |
|---|---|---|---|
| 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 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
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:
Browse additional application references in our FLAG® Literature portal.
- 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.
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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Description
Pricing
Storage Class
12 - Non Combustible Liquids
wgk
nwg
flash_point_f
Not applicable
flash_point_c
Not applicable
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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
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
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
Aleksandra Pajak et al.
PLoS genetics, 15(7), e1008240-e1008240 (2019-08-01)
The RNA helicase SUV3 and the polynucleotide phosphorylase PNPase are involved in the degradation of mitochondrial mRNAs but their roles in vivo are not fully understood. Additionally, upstream processes, such as transcript maturation, have been linked to some of these
Articles
Comparison of elution techniques for small-scale protein purification of FLAG® tag proteins using anti-FLAG® M2 magnetic beads.
Related Content
Protein and nucleic acid interaction reagents and resources for investing protein-RNA, protein-DNA, and protein-protein interactions and associated applications.
Réactifs pour interactions entre protéines et acides nucléiques, ressources permettant d'étudier les interactions protéines/ARN, protéines/ADN et protéines/protéines, et applications associées.
EZviewTM Red Protein A and ANTI-FLAG® M2 Affinity Gels: Immunoprecipitation with Enhanced Visibility Affinity Beads - Technical Article - July 2001
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What is the recommended short-term storage condition for FLAG M2 antibody — is 4°C acceptable?
1 answer-
Helpful?
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What is the recommended dilution for immunocytochemistry for this product ?
1 answer-
This product has not been validated for immunocytochemistry applications.
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Is there a recommended dilution for capture (sandwich) based ELISA?
1 answer-
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_coatingHelpful?
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What is the recommended dilution for conducting western blotting using this antibody?
1 answer-
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.pdfHelpful?
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What is the difference between F1804 and F3165 product?
1 answer-
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.
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What is the recommended dilution for conducting immunoprecipitation (IP) using the M2 antibody?
1 answer-
The general starting recommendation for conducting immunoprecipitation (IP) using the F3165, anti-FLAG M2 antibody, is 10 μg/mL.
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What type of light chain is present on the FLAG M2 antibody?
1 answer-
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?
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