GFP-Trap® M

Details for Product No. ABIN509401
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Target Name (Antigen)
Aequorea victoria
Magnetic Particles
Protein Complex Immunoprecipitation (Co-IP), Mass Spectrometry (MS), Enzyme Activity Assay (EAA), Affinity Measurement (AM), Chromatin Immunoprecipitation (ChIP), Pull-Down Assay (Pull-Down), Purification (Purif), Immunoprecipitation (IP)
Pubmed 24 references available
Quantity 20 tests
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Availability Will be delivered in 4 to 5 Business Days
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Catalog No. ABIN509401
442.20 $
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Purpose GFP-Trap® is a high quality GFP-binding protein coupled to a monovalent matrix (magnetic particles) for biochemical analysis of GFP fusion proteins and their interacting partners.
Brand GFP-Trap®
Sample Type Cell Extracts
Fragment heavy chain antibody (hcAb)
Specificity Binding capacity: 10 µL GFP-Trap®_M slurry binds 0.25 – 0.5 µg of GFP
Cross-Reactivity (Details) GFP-Trap® specifically binds to eGFP, wtGFP, GFP S65T, TagGFP, eYFP, YFP, Venus, Citrin, CFP. No binding to proteins derived from DsRed, all RFPs and TurboGFP can be detected.
Characteristics Antibodies – extremely powerful tools in biomedical research – are large complex molecules (~ 150 kDa) consisting of two heavy and two light chains. Due to their complex structure, the use of antibodies is often limited and hindered by batch-to-batch variations.

Camelidae (camels, dromedaries, llamas and alpacas) possess functional antibodies devoid of light chains, so-called heavy chain antibodies (hcAbs). hcAbs recognize and bind their antigens via a single variable domain (VHH). These VHH domains are the smallest intact antigen binding fragments (~ 13 kDa).

Nano-Traps are based on single domain antibody fragments (VHHs) derived from alpaca.
Components GFP-Trap® coupled to magnetic particles
Material not included Lysis buffer (CoIP), 10x RIPA buffer, Dilution buffer, Wash buffer, Elution buffer
Alternative Name GFP
Background The green fluorescent protein (GFP) and variants thereof are widely used to study the subcellular localization and dynamics of proteins. GFP fusion proteins can be expressed in different cell types at different expression levels by transient or stable transfection. Transient expression may provide quick informative results, however, in many cases it is necessary to generate stable cell lines that express the GFP fusion protein of interest at a level similar to the one of the endogenous protein. Quantification of GFP fusion proteins in cells can be tricky since existing methods, like fluorescence microscopy or Western Blotting, are often shows insufficient signal to noise ratios or high signal variabilities .
Research Area Tags/Labels
Application Notes Green fluorescent proteins (GFP) and variants thereof are widely used to study protein localization and dynamics. For biochemical analyses including mass spectroscopy and enzyme activity measurements these GFP-fusion proteins and their interacting factors can be isolated fast and efficiently (one step) via Immunoprecipitation using the GFP-Trap®. The GFP-Trap®_A enables purification of any protein of interest fused to GFP.

Bead size 0.5 - 1 µm

  • Robust and versatile tool for biochemical analyses of GFP-fusion proteins
  • Short incubation times (5 – 30 min)
  • Quantitative isolation of fusion proteins and transiently bound factors from cell extracts or organelles
  • Low unspecific binding
  • No contaminating heavy and light chains of conventional antibodies
  • Applicable in Chromatin Immunoprecipitation (ChIP)
Reagent Preparation Suggested buffer composition

  • Lysis buffer (CoIP): 10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA,0.5% NP-40
  • 10x RIPA buffer: 10 mM Tris/Cl pH 7.5, 150 mM NaCl, 5 mM EDTA, 0.1% SDS, 1% Triton X-100, 1% Deoxycholate
  • Dilution buffer: 10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA
  • Wash buffer: 10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA
  • Elution buffer: 200 mM glycine pH 2.5
Assay Procedure Before you start: Add 1ml PBS to your cells and scrape them off the petri dish.Transfer to precooled tube, spin 3 min at 500 x g and discard supernatant. Wash cell pellet twice with ice cold PBS, briefly resuspending the cells.
  • 1. For one immunoprecipitation reaction resuspend cell pellet (~10^7 mammalian cells) in 200 µL lysis buffer by pipetting (or using a syringe).
    optional: add 1 mM PMSF and Protease inhibitor cocktail (not included) to lysis buffer
    optional for nuclear/chromatin proteins: add 1 mg/ml DNase and 2.5 mM MgCl2 (not included) to lysis buffer
  • 2. Place the tube on ice for 30 min with extensively pipetting every 10 min.
  • 3. Spin cell lysate at 20.000x g for 5 -10 minutes at 4°C.
  • 4. Transfer supernatant to a pre-cooled tube. Adjust volume with dilution buffer to 500 µL – 1000 µL. Discard pellet.
    optional: add 1 mM PMSF and Protease inhibitor cocktail (not included) to dilution buffer
    note: the cell lysate can be frozen at this point for long-term storage at -80°C. For immunoblot analysis dilute 50 µL cell lysate with 50 µL 2x SDS-sample buffer(à refer to as input).
  • 5. Equilibrate GFP-Trap®_M beads in dilution buffer. Resuspend magnetic beads by vortexing and transfer 20 - 30 µL bead slurry in 500 µL ice cold dilution buffer. Magnetically separate beads until supernatant is clear. Discard supernatant and wash beads 2 more times with 500 µL ice cold dilution buffer.
  • 6. Add cell lysate to equilibrated GFP-Trap®_M beads and incubate the GFPTrap®_M beads with the cell lysate under constant mixing for 10 min – 2 h at room temperature or 4°C.
    note: during incubation of protein sample with the GFP-Trap®_M the final concentration of detergents should not exceed 0.2% to avoid unspecific binding to the matrix.
  • 7. Magnetically separate beads until supernatant is clear. For western blot analysis dilute 50 µL supernatant with 50 µL 2x SDS-sample buffer (à refer to as nonbound). Discard remaining supernatant.
  • 8. Wash beads three times with 500 µL ice cold wash buffer. After the last wash step, transfer beads to new tube.
    optional: increase salt concentration in the second washing step up to 500 mM
  • 9. Resuspend GFP-Trap®_M beads in 100 µL 2x SDS-Sample buffer or go to step 11.
  • 10. Boil resuspended beads for 10 minutes at 95°C to dissociate the immunocomplexes from the beads. The beads can be magnetically separated and SDS-PAGE is performed with the supernatant (à refer to as bound).
  • 11. optional: elute bound proteins by adding 50 µL 0.2 M glycine pH 2.5 (incubation time: 30 sec under constant mixing) followed by centrifugation. Transfer the supernatant to a fresh cup and add 5 µL 1M Tris base (pH 10.4) for neutralization. To increase elution efficiency this step can be repeated.
Restrictions For Research Use only
Concentration 500 µL resin
Buffer 1 x PBS,0.01% Sodium azide
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze.
Storage 4 °C
Expiry Date 12 months
Supplier Images
GFP-Trap® M Left (IP): Pulldown of GFP with GFP-Trap®_A and GFP-Trap®_M from 293T cell extracts. Input (I) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining. Right (Co-IP): Pulldown of GFP-PCNA with GFP-Trap®_A and GFP-Trap®_M from 293T cell extracts. Other bands: potential interaction partners of PCNA.
Product cited in: Mitchell, Lau, Lambert et al.: "Regulation of septin dynamics by the Saccharomyces cerevisiae lysine acetyltransferase NuA4." in: PLoS ONE, Vol. 6, Issue 10, pp. e25336, 2011 (PubMed).

Jovic, Kean, Szentpetery et al.: "TWO PI 4-KINASES CONTROL LYSOSOMAL DELIVERY OF THE GAUCHER DISEASE ENZYME, β-GLUCOCEREBROSIDASE." in: Molecular biology of the cell, 2012 (PubMed).

Metzger, Gache, Xu et al.: "MAP and kinesin-dependent nuclear positioning is required for skeletal muscle function." in: Nature, Vol. 484, Issue 7392, pp. 120-4, 2012 (PubMed).

Nakatogawa, Ohbayashi, Sakoh-Nakatogawa et al.: "The Autophagy-related Protein Kinase Atg1 Interacts with the Ubiquitin-like Protein Atg8 via the Atg8 Family Interacting Motif to Facilitate Autophagosome Formation." in: The Journal of biological chemistry, Vol. 287, Issue 34, pp. 28503-7, 2012 (PubMed).

Watanabe, Kobayashi, Yamamoto et al.: "Structure-based analyses reveal distinct binding sites for Atg2 and phosphoinositides in Atg18." in: The Journal of biological chemistry, 2012 (PubMed).

Kjær, Linch, Purkiss et al.: "Adenosine-binding motif mimicry and cellular effects of a thieno[2,3-d]pyrimidine-based chemical inhibitor of atypical protein kinase C isoenzymes." in: The Biochemical journal, Vol. 451, Issue 2, pp. 329-42, 2013 (PubMed).

Katzemich, Liao, Czerniecki et al.: "Alp/Enigma family proteins cooperate in Z-disc formation and myofibril assembly." in: PLoS genetics, Vol. 9, Issue 3, pp. e1003342, 2013 (PubMed).

Hsieh, Cheng, Lin: "Functional characterization of an abiotic stress-inducible transcription factor AtERF53 in Arabidopsis thaliana." in: Plant molecular biology, 2013 (PubMed).

Cheng, Tsai, Huang et al.: "Ser/Thr Kinase-Like Protein of Nicotiana benthamiana Is Involved in the Cell-to-Cell Movement of Bamboo mosaic virus." in: PLoS ONE, Vol. 8, Issue 4, pp. e62907, 2013 (PubMed).

Walte, Rüben, Birner-Gruenberger et al.: "Mechanism of dual specificity kinase activity of DYRK1A." in: The FEBS journal, Vol. 280, Issue 18, pp. 4495-511, 2013 (PubMed).

Gumy, Chew, Tortosa et al.: "The kinesin-2 family member KIF3C regulates microtubule dynamics and is required for axon growth and regeneration." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 33, Issue 28, pp. 11329-45, 2013 (PubMed).

Park, Sharma, Lefebvre et al.: "The endoplasmic reticulum-quality control component SDF2 is essential for XA21-mediated immunity in rice." in: Plant science : an international journal of experimental plant biology, Vol. 210, pp. 53-60, 2013 (PubMed).

Moser, Bensaddek, Ortmann et al.: "PHD1 links cell-cycle progression to oxygen sensing through hydroxylation of the centrosomal protein Cep192." in: Developmental cell, Vol. 26, Issue 4, pp. 381-92, 2013 (PubMed).

Kubacka, Kamenska, Broomhead et al.: "Investigating the consequences of eIF4E2 (4EHP) interaction with 4E-transporter on its cellular distribution in HeLa cells." in: PLoS ONE, Vol. 8, Issue 8, pp. e72761, 2013 (PubMed).

Linch, Sanz-Garcia, Soriano et al.: "A cancer-associated mutation in atypical protein kinase Cι occurs in a substrate-specific recruitment motif." in: Science signaling, Vol. 6, Issue 293, pp. ra82, 2013 (PubMed).

Pizon, Rybina, Gerbal et al.: "MURF2B, a Novel LC3-Binding Protein, Participates with MURF2A in the Switch between Autophagy and Ubiquitin Proteasome System during Differentiation of C2C12 Muscle Cells." in: PLoS ONE, Vol. 8, Issue 10, pp. e76140, 2013 (PubMed).

Kamenska, Lu, Kubacka et al.: "Human 4E-T represses translation of bound mRNAs and enhances microRNA-mediated silencing." in: Nucleic acids research, 2013 (PubMed).

Schneider, Steinberger, Strissel et al.: "The Arabidopsis TELLURITE RESISTANCE C Protein together with ALB3 is involved in Photosystem II Protein Synthesis." in: The Plant journal : for cell and molecular biology, 2014 (PubMed).

Ustün, König, Guttman et al.: "HopZ4 from Pseudomonas syringae, a member of the HopZ type III effector family from the YopJ superfamily, inhibits the proteasome in plants." in: Molecular plant-microbe interactions : MPMI, 2014 (PubMed).

Sharma, LeVaillant, Plant et al.: "Changes in expression of Class 3 Semaphorins and their receptors during development of the rat retina and superior colliculus." in: BMC developmental biology, Vol. 14, pp. 34, 2014 (PubMed).

General Rothbauer, Zolghadr, Tillib et al.: "Targeting and tracing antigens in live cells with fluorescent nanobodies." in: Nature methods, Vol. 3, Issue 11, pp. 887-9, 2006 (PubMed).

Agarwal, Hardt, Brero et al.: "MeCP2 interacts with HP1 and modulates its heterochromatin association during myogenic differentiation." in: Nucleic acids research, Vol. 35, Issue 16, pp. 5402-8, 2007 (PubMed).

Rothbauer, Zolghadr, Muyldermans et al.: "A versatile nanotrap for biochemical and functional studies with fluorescent fusion proteins." in: Molecular & cellular proteomics : MCP, Vol. 7, Issue 2, pp. 282-9, 2008 (PubMed).

Trinkle-Mulcahy, Boulon, Lam et al.: "Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes." in: The Journal of cell biology, Vol. 183, Issue 2, pp. 223-39, 2008 (PubMed).

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