A2220

Millipore

ANTI-FLAG® M2 Affinity Gel

purified immunoglobulin, buffered aqueous glycerol solution

Sinónimos:
Anti-ddddk, Anti-dykddddk, Monoclonal ANTI-FLAG® M2 antibody produced in mouse, ANTI-FLAG® M2 Affinity Agarose Gel
NACRES:
NA.32

Nivel de calidad

200

forma del anticuerpo

purified immunoglobulin

antibody product type

primary antibodies

clon

M2, monoclonal

formulario

buffered aqueous glycerol solution

analyte chemical class(es)

proteins (FLAG® tag, 3x FLAG®, DYKDDDDK tag)

aplicaciones

affinity chromatography: suitable (FLAG® peptide, Glycine, pH3.5, 3x FLAG® peptide)
immunoprecipitation (IP): suitable

matriz

(4% agarose bead; 45-165μm bead size)

isotipo

IgG1

capacidad

>0.6 mg/mL, resin binding capacity (FLAG-BAP)

conjugado

agarose conjugate

enviado en

wet ice

temp. de almacenamiento

−20°C

¿Está buscando productos similares? Visit Guía de comparación de productos

Descripción general

Anti-FLAG M2 Affinity gel is a mouse monoclonal antibody that is covalently attached to agarose. The antibody binds FLAG at the N-terminal, Met-N-terminal, C-terminal and internal locations of fusion proteins. Binding is calcium-independent.

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

Inmunógeno

DYKDDDDK

Aplicación

Anti-FLAG® M2 affinity gel has been used for western blotting, immunoprecipitation and for the purification of FLAG fusion proteins.

Browse additional application references in our FLAG® Literature portal.

Forma física

Suspension in buffered saline containing azide as preservative and 50% glycerol

Información legal

ANTI-FLAG is a registered trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Sigma-Aldrich Co. LLC

storage_class_code

10 - Combustible liquids

WGK Alemania

WGK 1

Punto de inflamabilidad F

Not applicable

Punto de inflamabilidad C

Not applicable

Certificado de Análisis

Certificado de origen

  1. Is my lysis buffer compatible with Product No. A2220, ANTI-FLAG® M2 Affinity Gel?

    The A2220 product information sheet (under Documents, above) contains a reagent compatibility table located on page 6. We do not recommend addition of SDS or reducing agents such as DTT, DTE or 2-mercaptoethanol to the resin. If you have a detergent listed in the table in a higher than recommended concentration, we recommend trying to dilute the sample before applying to the resin.

  2. How should I elute my protein when using Product No. A2220, ANTI-FLAG® M2 Affinity Gel?

    Elution with the peptide is the most gentle method. Acid elution (0.1 M glycine-HCL pH 3.5) is a more stringent method of elution, and should be evaluated for its effect on your protein if it is to be used in downstream applications. Boiling the resin in sample buffer is the most denaturing condition. If this condition is used, the resin cannot be re-used, due to the presence of SDS and/or reducing agents. The elution information can be viewed on A2220 product information sheet (under Documents, above).

  3. When using Product No. A2220, ANTI-FLAG® M2 Affinity Gel, I see bands at 20-25 kDa and 50-60 kDa appearing in my Westerns that are not my FLAG®-tagged protein. How can I prevent this?

    As a result of the conjugation, there may be some M2 antibody that is not conjugated to the resin, but is associated with the resin and may appear in acid elutions as heavy and light chain when using the anti-mouse IgG conjugated secondary antibody. We recommend an acid wash (0.1 M glycine-HCL pH 3.5) and neutralization of the resin (do not allow the acid wash to sit on the resin longer than 20 minutes) prior to applying the lysate.  Another way to avoid this is to use a directly conjugated FLAG® antibody for detection such as product A8592 ANTI-FLAG® M2 HRP, or the rabbit anti-FLAG® polyclonal antibody, F7425.

  4. I am using Product No. A2220, ANTI-FLAG® M2 Affinity Gel, and have a lot of non-specific proteins that are eluting with my FLAG®-tagged protein.  How can I get rid of these?

    The product bulletin for Product A2220, ANTI-FLAG® M2 affinity gel indicates:Pre-clear lysate with Mouse IgG-Agarose (Product A0919) to remove nonspecific binding proteins. Alternatively, you can use the unconjugated resin (Product 4B200) for this purpose. Other methods to remove non-specific binding from the resin would be to increase the stringency of the washes by increasing salt concentration (the resin can tolerate up to 1M NaCl) or including detergents that are compatible with the resin.

  5. When using ANTI-FLAG M2 Affinity Gel, Product A2220, should I use a 3X FLAG peptide or a 1X FLAG peptide to elute my protein?

    If you have a 3X FLAG-tagged protein, then you will need to use the 3X FLAG peptide.  If you have a 1X FLAG-tagged protein, you can use the 1X FLAG peptide or the 3X FLAG peptide.  We have not noticed a significant  difference in elution efficiency by using a 3X FLAG peptide on a 1X FLAG-tagged protein. 

  6. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  9. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Cédric Romilly et al.
Proceedings of the National Academy of Sciences of the United States of America, 116(32), 15901-15906 (2019-07-20)
In bacteria, stable RNA structures that sequester ribosome-binding sites (RBS) impair translation initiation, and thus protein output. In some cases, ribosome standby can overcome inhibition by structure: 30S subunits bind sequence-nonspecifically to a single-stranded region and, on breathing of the...
Yu Ti Cheng et al.
Proceedings of the National Academy of Sciences of the United States of America, 108(35), 14694-14699 (2011-08-30)
The nucleotide-binding domain and leucine-rich repeats containing proteins (NLRs) serve as immune receptors in both plants and animals. Overaccumulation of NLRs often leads to autoimmune responses, suggesting that the levels of these immune receptors must be tightly controlled. However, the...
Joshua M Baughman et al.
Nature, 476(7360), 341-345 (2011-06-21)
Mitochondria from diverse organisms are capable of transporting large amounts of Ca(2+) via a ruthenium-red-sensitive, membrane-potential-dependent mechanism called the uniporter. Although the uniporter's biophysical properties have been studied extensively, its molecular composition remains elusive. We recently used comparative proteomics to...
Hitoshi Nishizawa et al.
The Journal of biological chemistry, 279(19), 19391-19395 (2004-03-27)
Skeletal muscle is involved in the homeostasis of glucose and lipid metabolism. We hypothesized that the skeletal muscle produces and secretes bioactive factor(s), similar to adipocytokines secreted by fat tissue. Here, we report the identification of a novel secretory factor...
Nora Nonne et al.
Nucleic acids research, 38(4), e20-e20 (2009-12-04)
MicroRNAs (miRNAs) bind to Argonaute proteins, and together they form the RISC complex and regulate target mRNA translation and/or stability. Identification of mRNA targets is key to deciphering the physiological functions and mode of action of miRNAs. In mammals, miRNAs...
Artículos
The FLAG® Expression System is a proven method to express, purify and detect recombinant fusion proteins. Sigma®, the proven provider of FLAG®, now offers a magnetic bead for immunoprecipitation, protein purification, and the study of protein-protein interactions. The ANTI-FLAG® M2 Magnetic Bead is composed of murine derived, anti-FLAG® M2 monoclonal antibody attached to superparamagnetic iron impregated 4% agarose beads, with an average diameter of 50 µm. The M2 antibody is capable of binding to fusion proteins containing a FLAG peptide sequence at the N-terminus, Met-N-terminus, or C-terminus locations in mammalian, bacterial, and plant extracts.
Más información
Protocolos
Protocol for immunoprecipitation (IP) of FLAG fusion proteins using M2 monoclonal antibody 4% agarose affinity gels
Más información
Contenido relacionado
Protein purification techniques, reagents, and protocols for purifying recombinant proteins using methods including, ion-exchange, size-exclusion, and protein affinity chromatography.
Más información

Nuestro equipo de científicos tiene experiencia en todas las áreas de investigación: Ciencias de la vida, Ciencia de los materiales, Síntesis química, Cromatografía, Analítica y muchas otras.

Póngase en contacto con el Servicio técnico