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.
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):
affinity chromatography: suitable, immunoprecipitation (IP): suitable
Application:
IP, affinity chromatography
Citations:
204
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Unterstützung erhaltenQuality Level
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
1 of 4
Dieser Artikel | |||
|---|---|---|---|
| clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal | clone M2, monoclonal |
| shipped in wet ice | shipped in wet ice | shipped in wet ice | shipped in wet ice |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°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) - |
| isotype IgG1 | isotype IgG1 | isotype IgG1 | isotype IgG1 |
| analyte chemical class(es) proteins | analyte chemical class(es) proteins | analyte chemical class(es) proteins | analyte chemical class(es) - |
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.
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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Lagerklasse
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flash_point_f
Not applicable
flash_point_c
Not applicable
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Kara R Jones et al.
Molecular cancer therapeutics, 4(10), 1541-1547 (2005-10-18)
Incomplete DNA repair or misrepair can contribute to the cytotoxicity of DNA double-strand breaks. Consequently, interference with double-strand break repair, by pharmacologic or genetic means, is likely to sensitize tumor cells to ionizing radiation. The current studies were designed to
Matthew S Walters et al.
Journal of virology, 84(13), 6861-6865 (2010-04-16)
Varicella zoster virus encodes an immediate-early (IE) protein termed ORF61p that is orthologous to the herpes simplex virus IE protein ICP0. Although these proteins share several functional properties, ORF61p does not fully substitute for ICP0. The greatest region of similarity
Jiahai Zhou et al.
Proceedings of the National Academy of Sciences of the United States of America, 103(39), 14343-14348 (2006-09-16)
The unfolded protein response (UPR) is an evolutionarily conserved mechanism by which all eukaryotic cells adapt to the accumulation of unfolded proteins in the endoplasmic reticulum (ER). Inositol-requiring kinase 1 (IRE1) and PKR-related ER kinase (PERK) are two type I
Matthew Frieman et al.
Journal of virology, 83(13), 6689-6705 (2009-04-17)
The outcome of a viral infection is regulated in part by the complex coordination of viral and host interactions that compete for the control and optimization of virus replication. Severe acute respiratory syndrome coronavirus (SARS-CoV) intimately engages and regulates the
Ting-Ting Qu et al.
BMC cancer, 18(1), 27-27 (2018-01-06)
Lin28B and its paralog Lin28A are small RNA binding proteins that have similar inhibitory effects, although they target separate steps in the maturation of let-7 miRNAs in mammalian cells. Because Lin28B participates in the promotion and development of tumors mostly
Protokolle
Protokoll für die Immunpräzipitation (IP) von FLAG-Fusionsproteinen unter Einsatz des monoklonalen M2-Antikörper-Affinitätsgels (4 % Agarose)
Protocol for immunoprecipitation (IP) of FLAG fusion proteins using M2 monoclonal antibody 4% agarose affinity gels
Verwandter Inhalt
Protein expression technologies for various expression systems supporting research, therapeutics, and vaccine production.
Protein purification techniques, reagents, and protocols for purifying recombinant proteins using methods including, ion-exchange, size-exclusion, and protein affinity chromatography.
Proteinexpressionstechnologien für verschiedene Expressionssysteme zur Unterstützung von Forschung und der Herstellung von Arzneimitteln und Impfstoffen.
Proteinaufreinigungsverfahren, Reagenzien und Protokolle zur Aufreinigung rekombinanter Proteine unter Anwendung von Methoden wie Ionenaustausch, Größenausschluss und Proteinaffinitätschromatographie.
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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-
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What is the Department of Transportation shipping information for this product?
1 answer-
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.
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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-
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.
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When using Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, how can I elute my protein?
1 answer-
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.
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What is the binding capacity of the Product F2426, EZview™ Red ANTI-FLAG® M2 Affinity Gel clone M2, resin?
1 answer-
The binding capacity of the resin must be ? 0.6 mg/mL to meet specifications. This capacity will vary from lot to lot.
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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-
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.
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