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Human IgG2 FcSpyCatcher3

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Human IgG2-FcSpyCatcher3 thumbnail image 1 Human IgG2-FcSpyCatcher3 thumbnail image 2 Human IgG2-FcSpyCatcher3 thumbnail image 3
Human IgG2-FcSpyCatcher3 gallery image 1
Schematic image of Fab with SpyTag coupled to FcSpyCatcher3
Human IgG2-FcSpyCatcher3 gallery image 2
hIgG2-FcSpyCatcher3 (TZC016) was analyzed on a 4-20% Criterion TGX Stain-Free gel. Precision Plus Protein Unstained Protein Standard was loaded in lanes 1 & 4. Uncoupled hIgG2-FcSpyCatcher3 was run in lane 2 and hIgG2-FcSpyCatcher3 coupled to a SpyTagged antibody (AbD35759, Fab-F-Spy2-H format) in lane 3. Coupling was done with a 150% molar excess of Fab (25% excess of Fab over SpyCatcher sites) for 1 hr at RT. All lanes were loaded with 1 μg protein.
Human IgG2-FcSpyCatcher3 gallery image 3
Monodispersity assessment of hIgG2-FcSpyCatcher3 by SE-HPLC
Size exclusion chromatography (SEC) was performed on an analytical HPLC instrument. The peak corresponds to monomeric, fully assembled protein.
Column: MAbPac SEC-1.5 μm; dimension: 4.0 x 150 mm; mobile phase: 50 mM sodium phosphate, 200 mM arginine, 300 mM NaCl, pH 6.8, flow rate: 200 μl/min; inj. volume: 3 μl; temp.: 25 °C; detection: 280 nm; sample: 1 mg/ml.

Human IgG2-FcSpyCatcher3

Product Type
Recombinant Protein
Specificity
Human IgG2 FcSpyCatcher3

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Product Code Applications Pack Size List Price Your Price Qty
TZC016
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
IY 0.5 mg loader
List Price Your Price
loader

Human IgG2-FcSpyCatcher3 (hIgG2-FcSpyC3) contains a SpyCatcher3 (Keeble et al. 2019) fused to the hinge region, CH2 and CH3 of human IgG2 (Hentrich et al. 2020), resulting in a 76.4 kDa protein for the disulfide-bridged dimer. Each SpyCatcher3 subunit can form a stable covalent isopeptide bond with a second protein that includes a SpyTag. The reaction occurs spontaneously on mixing, is rapid, irreversible, high-yielding and shows good specificity. The reaction is robust at pH 5 to 8, at temperatures from +4°C to +37°C, in various buffer conditions (Ca2+/Mg2+ not needed) and in the presence of detergents. The reaction also occurs inside cells (in vivo). Since the speed of the coupling reaction is concentration dependent, it is recommended to use hIgG2-FcCatcher3 undiluted.

Human IgG2-FcSpyCatcher3 is compatible with Bio-Rad's recombinant HuCAL antibodies with a SpyTag2 at the C-terminus of the heavy chain.

Download FcSpyCatcher coupling protocol

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Target Species
Protein/peptide tag
Product Form
Purified recombinant protein - Liquid
Preparation
Recombinant protein expressed in mammalian cell line and purified by affinity chromatography
Source
HKB-11
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.1% ProClin 950
Purity
≥90% determined by SDS-PAGE under reducing conditions and visualized by coomassie blue staining
Approx. Protein Concentrations
80 µM (6.1 mg/ml)
Protein Molecular Weight
Predicted 76372 Da
Regulatory
For research purposes only
Guarantee
12 months from date of despatch
Acknowledgements
ProClin is a trademark of The Dow Chemical Company (“Dow”) or an affiliated company of Dow
This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
Licensed Use
For in vitro research purposes. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.

Store at -20°C only.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Immunoassay
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Instructions For Use
TZC016
View FcSpyCatcher Coupling Protocol

Description Product Code Applications Pack Size List Price Your Price Quantity
SpyTag3 Peptide BLP086 IY 1 mg loader
List Price Your Price
loader
Description SpyTag3 Peptide

Antibody Characterization Reference

  1. Hentrich, C. et al. (2020) Periplasmic Expression of SpyTagged Antibody Fragments Enables Rapid Modular Antibody Assembly
    [Epub ahead of print]
  2. Zakeri, B. et al. (2012) Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin.
    Proc Natl Acad Sci U S A. 109 (12): E690-7.
  3. Keeble, A.H. et al. (2019) Approaching infinite affinity through engineering of peptide-protein interaction.
    Proc Natl Acad Sci U S A. Dec 10 [Epub ahead of print].

TZC016

157513

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Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"

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