Antibody Panel to Green Fluorescent Protein (GFP) - FocusOn 118

Green Fluorescent Protein (GFP), due to the property of exhibiting bright green fluorescence when exposed to blue or UV light, has emerged as a powerful research tool for assessing gene expression and subcellular protein distribution in fixed or living cells. This feature makes it ideal as a marker for use in fluorescence microscopy, cytometry, tagging fusion proteins, and assaying transcriptional regulation from gene promoters in vivo.

Appealing in its ease of use, GFP folds into a chromophore without the need for accessory cofactors, enzymes or substrates other than molecular oxygen. Originally isolated from the jellyfish A. victoria, a number of laboratory mutations have been engineered into GFP to produce experimentally desirable variants with increased fluorescence, photostability, folding efficiency, and different excitation and emission peaks (blue, green, and yellow). The GFP gene or its variants are frequently attached to genes of interest using standard molecular and cell biology techniques to produce chimeric proteins, wherein the GFP serves as a reporter on the localization of the target protein. The small size and well-folded beta barrel structure make the highly fluorescing GFP relatively inert to the structure and properties of target proteins, and therefore ideal for this application. The GFP gene (as well as variants isolated from other marine species or engineered through mutations) has been introduced and expressed in a diverse set of model systems in bacteria, yeast and other fungi, fish, plants, insects, and mammalian cells and organisms.

Acris Antibodies offers a wide range of antibodies to detect GFP and its variants in various applications. The table below shows our recommended antibodies.

Confocal microscopy images of COS-7 cells transfected with expression constructs encoding membrane-tethered EGFP (membrane-EGFP; top) or nuclear Polycomb 2-EYFP fusion protein (Pc2-EYFP; bottom).

Fig. 1

Immunoprecipitation of GFP-NLS from HEK293 cells lysed in non-denaturating conditions using a rabbit anti-GFP antibody (lane 2) or a pre-immune rabbit serum (lane 3).

Fig. 2

Fig. 1: Confocal microscopy images of COS-7 cells transfected with expression constructs encoding membrane-tethered EGFP (membrane-EGFP; top) or nuclear Polycomb 2-EYFP fusion protein (Pc2-EYFP; bottom). The natural fluorescence of the produced proteins is shown in the green channel (left), the anti-GFP antibody Cat.-No. SP3005P signal was detected in the red channel (right). The system was carefully tested for overlap of these two optical channels and images were scanned separately in sequential scanning mode. The blue nuclear stain is also shown.

Fig. 2: Immunoprecipitation of GFP-NLS from HEK293 cells lysed in non-denaturating conditions using a rabbit anti-GFP antibody (lane 2) or a pre-immune rabbit serum (lane 3). Immunoprecipitates together with a sample of the cell lysate (lane 1) were separated on SDS-PAGE polyacrylamide gel and immunoblotted with the anti-GFP antibody Cat.-No. SP3005P.

Lineage tracing of EGFP expressing migrating pectoral girdle myogenic precursors in the chicken embryos via electroporation using TOL-2-GFP plasmid system.

Fig. 3

A GFP-tagged fusion protein was expressed in Nicotiana benthamiana and an IP with anti-GFP antibody Cat.-No. TP401 (0.25 µl from 1 mg/ml stock) was done from extracts.

Fig. 4

Fig. 3: Lineage tracing of EGFP expressing migrating pectoral girdle myogenic precursors in the chicken embryos via electroporation using TOL-2-GFP plasmid system. Vibratome sections stained with anti-GFP antibody Cat.-No. TP401 show the labeled muscle fiber soft hepectoral girdle in chicken embryos at stage HH27.
Data was kindly provided by: Nargis Khalida, Rizwan Rehimi: Institute of Anatomy, Dept. of Molecular Embryology, Ruhr University Bochum, Germany

Fig. 4: A GFP-tagged fusion protein was expressed in Nicotiana benthamiana and an IP with anti-GFP antibody Cat.-No. TP401 (0.25 µl from 1 mg/ml stock) was done from extracts. Western Blot analysis was performed using TP401 (1:5000).
Data was kindly provided by: Katharina Mueller, Georg Felix: Zentrum für Molekularbiologie der Pflanzen, Plant Biochemistry, University Tübingen, Germany

Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus in Mus musculus (coronal view, 20X magnification).

Fig. 5

Western blot of GFP recombinant protein detected with polyclonal anti-GFP antibody Cat.-No. R1091P (1 µg/ml).

Fig. 6

Fig. 5: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus in Mus musculus (coronal view, 20X magnification). Polyclonal goat anti-GFP Cat.-No. R1091P was used at 1:500 dilution in free floating immunohistochemistry to detect EGFP. Fluorchrome conjugated anti-goat IgG secondary antibody was used for detection at 1:500. Sections were counterstained with DAPI.
Source: Daniel A. Lee, Johns Hopkins School of Medicine, Baltimore, MD.

Fig. 6: Western blot of GFP recombinant protein detected with polyclonal anti-GFP antibody Cat.-No. R1091P (1 µg/ml). Lane 1 shows detection of a 33 kDa band corresponding to a GFP containing recombinant protein (arrowhead) expressed in HeLa cells. Lane 2 shows no staining of a mock transfected HeLa cell lysate.


Product Citations (Selection)

Catalog-No. Citations
R1091P 1
TP401 2
1
Yuko Suzuki et al.: Unique secretory dynamics of tissue plasminoge activator and its modulation by plasminogen activator inhibitor-1 in vascular endothelial cells.
Blood, Jan 2009; 113: 470-478. (Use of R1091T)
2
Markus Albert et al.: The Arabidopsis thaliana pattern recognition receptors for bacterial EF-TU and flagellin can be combined to form functional chimeric receptors.
J Biol Chem, Jun 2010; 285(25): 19035-19042.
2
Philipp Angenendt et al.: Generation of high density protein microarrays by cell-free in situ expression of unpurified PCR products.
Mol Cell Proteomics, Sep 2006; 5(9): 1658-1666.
2
Nan Li et al.: Regulation of neural crest cell fate by the retinoic acid and Pparg signalling pathways.
Development, Feb 2010; 137(3): 389-394.
2
Lionel Franz Poulin et al.: The dermis contains langerin+ dendritic cells that develop and function independently of epidermal Langerhans cells.
J Exp Med, Dec 2007; 204(13): 3119-3131.
2
Faisal Yusuf et al.: Inhibitors of CXCR4 affect the migration and fate of CXCR4+ progenitors in the developing limb of chick embryos.
Dev Dyn, Nov 2006; 235(11): 3007-3015.
2
Stephanie Blachon et al.: Nucleo-cytoplasmic shuttling of high risk human papillomavirus E2 proteins induces apoptosis.
J Biol Chem, Oct 2005; 280(43): 36088-36098.
2
Kerstin Laib Sampaio et al: Human cytomegalovirus labeled with green fluorescent protein for live analysis of intracellular particle movements.
J Virol, Mar 2005, 79(5): 2754-2767.

Remarks

  • TP401 has also been described to work in IP and in filter binding assays.

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

GFP antibody

Figure 1. Confocal microscopy images of COS-7 cells transfected with expression constructs encoding membrane-tethered EGFP (membrane-EGFP; top) or nuclear Polycomb 2-EYFP fusion protein (Pc2-EYFP; bottom). The natural fluorescence of the produced proteins is shown in the green channel (left), the anti-GFP antibody signal was detected in the red channel (right). The system was carefully tested for overlap of these two optical channels and images were scanned separately in sequential scanning mode. The blue nuclear stain is also shown. Rabbit Purified All Species ICC/IF, IP, WB
0.1 mg / €230.00
  Acris Antibodies GmbH

GFP antibody

Lineage tracing of EGFP expressing migrating pectoral girdle myogenic precursors in the chicken embryos via electroporation using TOL-2-GFP plasmid system. Vibratome sections stained with anti-GFP antibody Cat.-No. TP401 show the labeled muscle fiber soft hepectoral girdle in chicken embryos at stage HH27.
Data was kindly provided by: Nargis Khalida, Rizwan Rehimi: Institute of Anatomy, Dept. of Molecular Embryology, Ruhr University Bochum, Germany Rabbit Purified A. victoria E, IP, WB
0.2 mg / €290.00
  Torrey Pines Biolabs

GFP antibody

GFP Goat AP A. victoria C, E, WB
1 mg / €620.00
  Acris Antibodies GmbH

GFP (Ads. to Hu, Ms, Rt Serum Proteins) antibody

Polyclonal anti-GFP antibody at a 1/1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nervous system. Fluorochrome conjugated anti-Goat secondary antibody was used for detection at 1/300. Panel A shows a lateral view (ventral left). Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up.
Personal Communication, Helmata Mistry, Washington University School of Medicine, St. Louis, MO. Goat Biotin A. victoria C, E, ICC/IF, WB
1 mg / €510.00
  Acris Antibodies GmbH

GFP antibody

GFP Goat FITC A. victoria C, E, ICC/IF, WB
1 mg / €510.00
  Acris Antibodies GmbH

GFP (Ads. to Hu, Ms, Rt Serum Proteins) antibody

GFP Goat HRP C, E, WB
1 mg / €510.00
  Acris Antibodies GmbH

GFP (Ads. to Hu, Ms, Rt Serum Proteins) antibody

Immunofluorescence Microscopy of Anti-GFP (GOAT) Antibody: Tissue: E5.5 Hex-GFP transgenic mouse embryo. Primary antibody: Goat anti-GFP was used at 1/500 dilution. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody at 1/10,000 for 45 min at RT. Staining: GFP as green fluorescent signal with DAPI blue counterstain. Goat Aff - Purified C, E, ICC/IF, WB
1 mg / €490.00
  Acris Antibodies GmbH

GFP antibody

GFP Goat TRITC E, ICC/IF, WB
1 mg / €510.00
  Acris Antibodies GmbH

GFP antibody

GFP Goat Texas Red C, ICC/IF
1 mg / €510.00
  Acris Antibodies GmbH

GFP antibody

Western Blot of Anti-GFP (MOUSE) Monoclonal Antibody Peroxidase Conjugate. Lane 1: 50ng of GFP. Lane 2: none. Primary antibody: none. Secondary antibody: Anti-GFP (Mouse) Monoclonal Antibody Peroxidase Conjugate (Cat.-No R1461HRP) secondary antibody was used at 1:1000 for 45 min at RT. Block: 1% BSA-TTBS (p/n MB-013, diluted to 1X) 30 min at 20°C. Predicted/Observed size: 28 kDa for GFP. Other band(s): none. Mouse IgG2a 9F9.F9 HRP A. victoria C, E, WB
1 mg / €720.00
  Acris Antibodies GmbH

GFP antibody

GFP Mouse IgG2a 9F9.F9 AP A. victoria C, E, WB
1 mg / €830.00
  Acris Antibodies GmbH

GFP antibody

GFP Mouse IgG2a 9F9.F9 Biotin A. victoria C, E, WB
1 mg / €720.00
  Acris Antibodies GmbH

GFP antibody

GFP Mouse IgG2a 9F9.F9 FITC A. victoria ICC/IF
1 mg / €720.00
  Acris Antibodies GmbH

GFP antibody

Western blot of GFP recombinant protein detected with monoclonal anti-GFP antibody. GFP recombinant protein was expressed in HeLa cells, where 50 ng (lane 1), 100 ng (lane 2) and 500 ng (lane 3) of lysate were loaded per lane. Anti-GFP detects a 27 kDa band corresponding to the epitope tag GFP. The cell lysates were prepared in a RIPA buffer containing 200 mM NaCl. A 4-12% Bis-Tris gradient gel was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking with 5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1.0 ?g/ml for 1 h at room temperature followed by washes and reaction with a 1:2,500 dilution of IRDye® 800 conjugated Goat-a-Mouse IgG [H&L]IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results. Mouse IgG2a 9F9.F9 Purified A. victoria C, E, WB
1 mg / €620.00
  Acris Antibodies GmbH

GFP antibody

GFP Mouse IgG2a 9F9.F9 Texas Red A. victoria E, ICC/IF, WB
1 mg / €720.00
  Acris Antibodies GmbH

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