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F2426

EZview™ Red ANTI-FLAG® M2-Affinitätsgel

clone M2

Synonym(e):

Monoklonaler ANTI-FLAG® M2-Antikörper in Maus hergestellte Antikörper, Anti-ddddk, Anti-dykddddk

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Über diesen Artikel

NACRES:
NA.32
UNSPSC Code:
12352203
Clone:
M2, monoclonal
Technique(s):
FLAG purification: suitable, affinity chromatography: suitable, immunoprecipitation (IP): suitable
Application:
FLAG purification, IP, affinity chromatography
Citations:
204
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Quality Segment

clone

M2, monoclonal

analyte chemical class(es)

proteins

technique(s)

FLAG purification: suitable, 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

Das EZview Red Anti-FLAG M2-Affinitätsgel ist ein Harz, das aus Anti-FLAG M2-Antikörpern besteht, die kovalent an Agarose-Beads (4 % Agarose) gebunden sind. Das Affinitätsgel wird verwendet, um FLAG-Fusionsproteine an Proben wie Zelllysate und Gewebe zu binden und FLAG-getaggte Proteine zur Vorbereitung für Immunpräzipitations-Assays aufzureinigen. Der rote Farbstoff sorgt für eine bessere Sichtbarkeit und effizientere Ergebnisse. Agarose-Beads binden an N-terminalen, Met-N-terminalen, C-terminalen FLAG-Fusionsproteinen und 3xFLAG-getaggten Fusionsproteinen.

Application

Immunpräzipitation (IP) von FLAG- und 3xFLAG-markierten Fusionsproteinen.

Elution – FLAG-Peptid, Glycin, pH 3,5, 3xFLAG-Peptid

Weitere Produktinformationen finden Sie in unserem FLAG® Literatur-Portal.

Biochem/physiol Actions

Geeignet für die Aufreinigung von N-terminalen, Met-N-terminalen, C-terminalen FLAG-Fusionsproteinen und 3xFLAG-Fusionsproteinen.

Physical form

1:1 (v/v) Suspension in PBS enthält 50 % Glycerin und 15 ppm 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

FLAG®-Affinitätsgele, FLAG®-Tag, 3xFLAG®-Tag, DYKDDDDK-Tag

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Dieser Artikel
M8823A4596E6779
description

clone M2

description

affinity isolated antibody

description

-

description

-

clone

M2, monoclonal

clone

M2, monoclonal

clone

-

clone

-

shipped in

wet ice

shipped in

wet ice

shipped in

-

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)

immunoprecipitation (IP): suitable, affinity chromatography: suitable

technique(s)

-

technique(s)

immunoprecipitation (IP): suitable, affinity chromatography: suitable

isotype

IgG1

isotype

IgG1

isotype

IgG12b

isotype

-

analyte chemical class(es)

proteins

analyte chemical class(es)

proteins

analyte chemical class(es)

-

analyte chemical class(es)

proteins (hemagglutinin)


Lagerklasse

10 - Combustible liquids

Flammpunkt (°F)

Not applicable

Flammpunkt (°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 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?

  3. 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?

  4. 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?

  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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