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Erstklassige Werkzeuge für die Immunpräzipitation und -aufreinigung

GFP Catcher - High-Affinity Single-Domain Antibodies

Julian Pampel

antibodies-online bietet mit den GFP-Catchern das ideale Werkzeug für den effektiven Pulldown von GFP-Fusionsproteinen. Unsere Catcher-Produktlinien basieren auf rekombinanten Single-Domain-Antikörpern (sdAbs), die von Lamas oder Alpakas stammen. Diese Art von Affinitätsmolekülen bietet erhebliche Vorteile gegenüber herkömmlichen IgG-Molekülen. Alle unsere Produkte werden intern hergestellt. Sie sind hochspezifisch und verfügen über Affinitäten, die auf ein breites Spektrum von Anwendungen zugeschnitten sind.

Was sind GFP-Catcher?

Der GFP-Catcher ist ein gebrauchsfertiges Affinitätsharz für die Immunpräzipitation von GFP-Fusionsproteinen. GFP-Catcher und Catcher im Allgemeinen basieren auf einem hochaffinen Single-Domain-Antikörper (sdAb), der kovalent auf 4 % vernetzten Agarose-Perlen immobilisiert ist. Der GFP-Catcher (ABIN5311508) zeichnet sich somit durch eine hohe Affinität und überlegene Kapazität für GFP-Fusionsproteine aus, während der unspezifische Hintergrund vernachlässigbar ist (Abb. 1). Due to the covalent and stable linkage of an GFP-specific sdAb to the agarose support, bound GFP fusion proteins are eluted from GFP Selector without contaminating IgG chains (* Band originating from maturation of GFP). GFP-Catchers immobilize a wide range of GFP derivatives: EGFP, mEGFP, Sirius, tSapphire, Cerulean, eCFP, mTurquoise, acGFP, Emerald, superecliptic pH luorin, paGFP, superfolder GFP, eYFP, mVenus and Citrine and most common CFP and YFP variants are being recognized. GFP-Catchers are compatible not only with physiological buffers but also with high stringency buffers. GFP-Catchers thus provide great freedom to adjust the binding and washing conditions to the experimental needs. In comparison with competitor products, our antibodies-online GFP Catcher (ABIN5311508) offers an increased binding capacity, 1 µl of packed beads is able to bin 4 μg GFP.

GFP Catcher ABIN5311508

Fig.1: (A) Pull-down of GFP from a mixture of GFP, mCherry and mTagBFP (B) Immunoprecipitation of GFP (arrow) from HeLa lysate. (C) Control experiment using functionalized beads lacking sdAbs.

Advantages of Catchers for IP

The Agarose beads of our catchers offer, in comparison with magnetic beads, the lowest background and high binding capacity for IP. The innovative, oriented and selective attachment via a flexible linker guarantees a high accessibility of the sdAbs and largely eliminates batch-to-batch variations. Due to the single-chain nature of sdAbs and their covalent attachment, no "leakage" of light and heavy chains from IgGs is observed during elution with SDS sample buffer. The absence of heavy & light antibody chains enables to drastically decrease the incubation time to 5-30 minutes. Even under harsh conditions the antibody binds with very high affinity and pulls out even low abundant proteins.

All Catchers at one glance

Produkt
Kat. Nr.
Reaktivität
Applikation
Sample Type
Menge
Datenblatt
Kat. Nr. ABIN5311510
Reaktivität Discosoma
Applikation Co-IP, IP, Purif, ChIP, RIP
Sample Type Cell Extracts
Menge 2000 μL
Kat. Nr. ABIN5311504
Reaktivität E. coli
Applikation Co-IP, IP, Purif, ChIP, RIP
Sample Type Cell Extracts
Menge 2000 μL
Kat. Nr. ABIN5311508
Reaktivität Aequorea victoria
Applikation Co-IP, IP, Purif, ChIP, RIP
Sample Type Cell Extracts
Menge 2000 μL
Kat. Nr. ABIN5311512
Reaktivität Entacmaea quadricolor
Applikation Co-IP, IP, Purif, ChIP, RIP
Sample Type Cell Extracts
Menge 2000 μL

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  • We offer Catchers as reliable replacement for Traps in IP experiments
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Catcher Applications

All catcher agarose beads are validated and tested for a variety of applications:

  • Protein Complex Immunoprecipitation (Co-IP)
  • Immunoprecipitation (IP)
  • Purification (Purif)
  • Chromatin Immunoprecipitation (ChIP)
  • RNA-Binding Protein Immunoprecipitation (RIP)

Additive Products

Catcher-Control is a control resin for all Catcher resins, which are based on 4 % cross-linked agarose beads. It was prepared using the same activation and quenching chemistry that was used for preparation of Catcher resins, but is not charged with any single-domain antibody. Catcher-Control is thus the ideal resin for pre-depletion of extracts or as a side-by-side specificity control for any Catcher resins. When using as a specificity control, Catcher-Control and the corresponding sdAb-charged Catcher Resin should be used under identical experimental conditions.

Protocols

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