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F2426

Gel d′affinità EZview rosso ANTI-FLAG® M2

clone M2

Sinonimo/i:

Anticorpo monoclonale ANTI-FLAG® M2, Anti-ddddk, Anti-dykddddk

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Informazioni su questo articolo

NACRES:
NA.32
UNSPSC Code:
12352203
Clone:
M2, monoclonal
Technique(s):
affinity chromatography: suitable, immunoprecipitation (IP): suitable
Application:
IP, affinity chromatography
Citations:
204
Prezzi e disponibilità al momento non sono disponibili
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Quality Segment

clone

M2, monoclonal

analyte chemical class(es)

proteins

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

matrix

4% agarose bead; 45-165μm bead size

isotype

IgG1

capacity

≥0.6 mg/mL, gel binding capacity

shipped in

wet ice

storage temp.

−20°C

General description

Il gel d′affinità EZ view rosso anti-FLAG M2 è una resina costituita dall′anticorpo anti-FLAG M2, legato con legame covalente a microsfere di agarosio al 4%. Il gel di affinità è usato per legare le proteine di fusione FLAG ai campioni, come lisati cellulari e tessuti, per purificare le proteine taggate con FLAG in preparazione ai saggi di immunoprecipitazione. Grazie al colorante rosso che migliora la visibilità si ottengono risultati più efficienti. Le microsfere di agarosio si legano alle proteine di fusione con FLAG N-terminali, Met-N-terminali e C-terminali, e alle proteine di fusione taggate con 3xFLAG.

Application

Immunoprecipitazione (IP) di proteine di fusione taggate FLAG e 3xFLAG.

Eluizione - peptide FLAG
, glicina, pH 3,5, peptide 3x FLAG
®Esplora tutti i dettagli di prodotto sul nostro portale dedicato alle applicazioni FLAG

Biochem/physiol Actions

Adatto a purificare proteine di fusione con FLAG N-terminali, Met-N-terminali, C-terminali, proteine di fusione 3xFLAG.

Physical form

Sospensione 1:1 (v/v) in PBS contenente glicerolo al 50% e 15 ppm di Kathon

Legal Information

ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
EZview is a trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Gel di affinità FLAG®, tag FLAG®, tag 3xFLAG®, tag DYKDDDDK

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Questo articolo
A2220M8823E6654
clone

M2, monoclonal

clone

M2, monoclonal

clone

M2, monoclonal

clone

-

shipped in

wet ice

shipped in

wet ice

shipped in

wet ice

shipped in

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

2-8°C

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

technique(s)

affinity chromatography: suitable, immunoprecipitation (IP): suitable

isotype

IgG1

isotype

IgG1

isotype

IgG1

isotype

-

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200


Classe di stoccaggio

10 - Combustible liquids

flash_point_f

Not applicable

flash_point_c

Not applicable



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Questions

1–6 of 6 Questions  
  1. When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, should I use a 3X FLAG peptide or a 1X FLAG peptide to elute my protein?

    1 answer
    1. 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.

      Helpful?

  2. When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, 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?

    1 answer
    1. 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 a 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 ant-FLAG M2 HRP, or the rabbit anti-FLAG polyclonal antibody, F7425.

      Helpful?

  3. When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, I have a lot of non-specific proteins that are eluting with my FLAG-tagged protein. How can I get rid of these?

    1 answer
    1. One way to remove non-specific proteins is to pre-bind the protein lysate with unconjugated resin.  We recommend product 4B200 for this purpose. Other methods 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.

      Helpful?

  4. What is the binding capacity of the Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, resin?

    1 answer
    1. The binding capacity of the resin must be   ? 0.6 mg/mL to meet specifications.  This capacity will vary from lot to lot.

      Helpful?

  5. When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, how can I elute my protein?

    1 answer
    1. 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.

      Helpful?

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

    1 answer
    1. 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.

      Helpful?

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