- Chromotek-GFP-Trap®
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Chromotek-GFP-Trap® is a high quality, single domain GFP-binding protein coupled to a monovalent matrix. It can be a robust tool for:
- Identify interacting proteins
- Measure enzyme activity
- Determine DNA or RNA binding
- Map DNA binding sites
- ChIP-chip assays
- CLIP assays (in vivo Cross-Linking and ImmunoPrecipitation)
Rothbauer U et al. A versatile nanotrap for bioc...(PubMed)
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- Pull Down with GFP-Trap® vs others
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Features:
- The isolation/depletion of GFP-fusion using GFP-Trap is quantitative.
- There are no IgG heavy or light chain backgrounds at all for subsequent Western blotting.
Let the Data Speak!
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- What Have Been Achieved
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Rogowski et al. 2009 Cell 137, 1076-1087
Abstract: Potential polyglycylation (addition of chains of glycine on the gamma-carboxyl groups of specific glutamate residues of target proteins) substrates were tested as EYFP-fusions after co-IP with Camelid antibody based GFP-Trap resins (also bind to GFP derivatives such as EYFP). True substrates were positively identified.
More References 
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- What Have Been Achieved
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Webby et al. 2009 Science 235, 90-93
Abstract: A posttranslational lysyl-hydroxylation enzyme was fused to GFP and isolated with GFP-Trap. The associated proteins were analyzed by mass spectrometry (MS), substrates in pre-mRNA splicing such as U2AF and other SR proteins identified, and the hypothesis that the enzyme, Jumonji-6 protein, is involved in alternative splicing was proven correct.
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- What Users Say:
- “Your GFP-Trap® is great! I have never seen such efficient reagent for pull down.
- Prof. Dr. Tomoyuki Tanaka, Wellcome Trust Centre for Gene Regulation & Expression, University of Dundee
- “We recently had excellent MS results with the GFP-Trap® "
- Dr. Gwyneth Ingram, IMPS, Edinburgh
- “…we were gob smacked when we did our own IP and saw the glowing green GFP-Trap® beads.."
- Prof. Dr. Laura Trinkle-Mulcahy, University of Ottawa
- More Testimonials
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- Immunoprecipitation (IP)
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Immunoprecipitation is a process of isolating a protein as an antigen by using antibodies against it. It is a powerful tool for studying proteins in biological samples and, in case of Co-IP (meaning immunoprecipitation of complexes containing a known antigen), for analyzing protein-protein interactions. Similar technologies such as chromatin immunoprecipitation (ChIP), RNA immunoprecipitation (RIP), or crosslinked and immunoprecipitation of RNA-protein complexes (CLIP) analysis of protein-DNA or protein-RNA interactions.
The major obstacle for achieving effective immunoprecipitation is ......
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- Choosing Tags for IP Assay
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Among the most commonly used tags are: FLAG, Myc, HA, V5, T7, and His, which are quite small in size and in theory less likely to interfere. GST and GFP are well documented to form self-contained and stable structures independent of their fusion partners and proven to not interfere in many cases despite their larger size (in between 20-30kD.) A top choice for pulldown experiments, GST can bind to glutathione beads directly. GFP or other FPs are excellent tags having both the advantages of being a visualization module to follow the protein both inside cells and during pulldown. ......
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- Choosing Lysis Buffers for IP Assay
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For immunoprecipitation experiments, how cells are lysed and proteins extracted from insoluble fractions is critical for the success of pull-down and subsequent assays. For certain types of enzymatic reactions, the lysis buffer needs to be devoid of reducing agents, strong detergents, high salt concentration, etc. There are less known detergents that have properties that may be more suitable for lysing the cells while maintaining certain levels of protein-protein interactions, as used in the AlleleExtract-M buffers.
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- AlleleExtract Lysis Buffers
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As a convenient starting solution, AlleleExtract-B Protein Reagent provides a single-step method of gently extracting proteins from a wide range of bacterial cells.
AlleleExtract-M Protein Reagent provides an effective and gentle method for extracting proteins from mammalian cells.
Allele´s 5X Insect Cell Lysis Buffer has a classic composition specifically formulated for lysing insect cells.
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- Anti-GFP Monoclonal Antibody
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Allele Biotech´s Anti-GFP monoclonal antibody provides a simple solution to detect the expression of a GFP-tagged protein in cells or immunoprecipitation assay.
This antibody is best used when combined with Allele´ GFP-Trap resins, which are immobilized camelid antibodies against GFP with much better precipitation efficiency than monoclonal or polyclonal antibodies. After pulldown, Allele monoclonal antibody agaisnt GFP can be used to quantitatively analyze the precipitation efficiency, as demonstrated in experiments related to Allele’s GFP-Trap
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- One-stop Pull Down Assay Services
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Immunoprecipitation is commonly used to isolate proteins from cell lysate. By analyzing factors that co-immunoprecipitate with the protein pulled down by the precipitating antibodies, researchers can study complex formation, protein-protein, protein-DNA, or protein-RNA interactions.
Cat #: ABP-CS-IMP0001
Description: Immunoprecipitation of fusion proteins with GFP (EGFP or any jelly fish GFP derivatives except for CFP). Analysis of precipitated proteins by SDS-PAGE and by Western blotting with anti-GFP monoclonal (ABG-MP-MMGFP10) or polyclonal antibodies (ABP-PAB-PAGFP10)
Required Materials: Pellet of 107 cells per precipitation
Deliverables: Precipitated proteins in 100ul 2X SDS loading buffer, or 55ul neutralized elution buffer (Glycine/Tris), SDS page image and Western blotting with anti-GFP antibodies
Duration: 3 days
Price (USD): Includes: GFP-Trap, anti-GFP antibodies, and Western detection reagents for STORM imaging.
$375 first sample
$675 two samples
$275 each additional sample for more than two samples
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