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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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.
biological source
mouse
Quality Segment
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
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
1 of 1
This Item | |||
|---|---|---|---|
| clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal |
| conjugate unconjugated | conjugate unconjugated | conjugate - | conjugate agarose conjugate |
| antibody form purified immunoglobulin (Purified IgG1 subclass) | antibody form affinity isolated antibody | antibody form - | antibody form purified immunoglobulin |
| species reactivity all | species reactivity all | species reactivity - | species reactivity - |
| biological source mouse | biological source mouse | biological source - | biological source - |
| form buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide) | form buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4) | form - | form buffered aqueous glycerol solution |
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Storage Class
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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What is the recommended short-term storage condition for FLAG M2 antibody — is 4°C acceptable?
1 answer-
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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.
Helpful?
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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.
Helpful?
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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.
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