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GFP

Principal name

GFP antibody

Selected pictures

SP3005P - 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.

SP3005P - Figure 2. Immunoprecipitation of GFP-NLS from HEK293 cells using anti-GFP antibody. HEK293 cells were transfected with expression construct encoding GFP-NLS protein. Twenty hours post transfection cells were lysed in non-denaturating conditions (Lysis buffer: 20 mM Tris, pH 7.5, 100 mM NaCl, 0.5% Triton X-100, inhibitors of proteases). Aliquots of cell lysate were immunoprecipitated 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. The positions of molecular weight markers in kDa are indicated at the left.

Gene ID

3020 (H3F3A)

Ncbi ID

6100, NP_002098

Available reactivities

All Species, A. victoria, Hu (Human), Hydra

Available hosts

Rabbit, Chicken, Goat, Mouse, Sheep

Available applications

Western blot / Immunoblot (WB), Frozen Sections (C), Immunoprecipitation (IP), Immunocytochemistry/Immunofluorescence (ICC/IF), Enzyme Immunoassay (E), Paraffin Sections (P), Flow Cytometry (F), Dot blot (Dot)

Selected product citations ‐ of TP401

1. Albert M, Jehle AK, Mueller K, Eisele C, Lipschis M, Felix G. Arabidopsis thaliana pattern recognition receptors for bacterial elongation factor Tu and flagellin can be combined to form functional chimeric receptors. J Biol Chem. 2010 Jun 18;285(25):19035-42. doi: 10.1074/jbc.M110.124800. Epub 2010 Apr 21. PubMed PMID: 20410299.
2. Angenendt P, Kreutzberger J, Glökler J, Hoheisel JD. Generation of high density protein microarrays by cell-free in situ expression of unpurified PCR products. Mol Cell Proteomics. 2006 Sep;5(9):1658-66. Epub 2006 Jul 5. PubMed PMID: 16825183.
3. Li N, Kelsh RN, Croucher P, Roehl HH. Regulation of neural crest cell fate by the retinoic acid and Pparg signalling pathways. Development. 2010 Feb;137(3):389-94. doi: 10.1242/dev.044164. PubMed PMID: 20081187.
4. Poulin LF, Henri S, de Bovis B, Devilard E, Kissenpfennig A, Malissen B. The dermis contains langerin+ dendritic cells that develop and function independently of epidermal Langerhans cells. J Exp Med. 2007 Dec 24;204(13):3119-31. Epub 2007 Dec 17. PubMed PMID: 18086861.
5. Yusuf F, Rehimi R, Moroşan-Puopolo G, Dai F, Zhang X, Brand-Saberi B. Inhibitors of CXCR4 affect the migration and fate of CXCR4+ progenitors in the developing limb of chick embryos. Dev Dyn. 2006 Nov;235(11):3007-15. PubMed PMID: 16958136.
6. Blachon S, Bellanger S, Demeret C, Thierry F. Nucleo-cytoplasmic shuttling of high risk human Papillomavirus E2 proteins induces apoptosis. J Biol Chem. 2005 Oct 28;280(43):36088-98. Epub 2005 Aug 31. PubMed PMID: 16135518.
7. Sampaio KL, Cavignac Y, Stierhof YD, Sinzger C. Human cytomegalovirus labeled with green fluorescent protein for live analysis of intracellular particle movements. J Virol. 2005 Mar;79(5):2754-67. PubMed PMID: 15708994.

Background of GFP antibody

Green fluorescence protein (GFP) is a 27 KDa protein derived from the bioluminiscent jellyfish Aquorea victoria, emiting green light (509 nm) when excited (excitation by Blue or UV light, absorption peak at 395 nm).
GFP is a useful tool in cell biology research, as its intrinsic fluorescence can be visualized in living cells. Light-stimulated GFP fluorescence is species-independent and a fluorescence has been reported from many different types of GFP-expressing hosts, including microbes, invertebrates, vertebrates and plants. No exogenous substrates and cofactors are required for the fluorescence of GFP, since GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein.
GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP is widely used as a reporter (tag) for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without any further staining. Other applications of GFP include measurement of distance between proteins through fluorescence energy transfer (FRET) protocols.
To increase a fluorescence intensity of GFP, chromophore mutations have been created. The Enhanced GFP has a fluorescence 35 times more intense than the wt-GFP. Mutagenesis of GFP has produced also many mutants (e.g. Yellow Fluorescent Protein, Cyan Fluorescent Protein) with warying spectral properties. Antibodies raised against full-length GFP variants should also detect other variants of the protein.

General readings

Schoch KG et al. A subset of mouse tracheal epithelial basal cells generates large colonies in vitro. Am J Physiol Lung Cell Mol Physiol 286:L631-42 (2004).
Karabay A. et al Axonal growth is sensitive to the levels of katanin, a protein that severs microtubules. The Journal of Neuroscience 24:5778-5788 (2004).Gongidi V et al. SPARC-like 1 Regulates the Terminal Phase of Radial Glia-Guided Migration in the Cerebral Cortex. Neuron 41:57-69 (2004).
Fischer AC et al. Successful transgene expression with serial doses of aerosolized rAAV2 vectors in rhesus macaques. Mol Ther 8:918-26 (2003).
Wong JM et al. Subnuclear shuttling of human telomerase induced by transformation and DNA damage. Nat Cell Biol 4:731-6 (2002).
Zhang QX et al. Identification of structurally important domains of lipid phosphate phosphatase-1: implications for its sites of action. Biochem J 345 Pt 2:181-4 (2000).
Mesaeli N et al. Calreticulin is essential for cardiac development. J Cell Biol 144:857-68 (1999).
McCabe JB & Berthiaume LG Functional roles for fatty acylated amino-terminal domains in subcellular localization. Mol Biol Cell 10:3771-86 (1999).
Jasinska R et al. Lipid phosphate phosphohydrolase-1 degrades exogenous glycerolipid and sphingolipid phosphate esters. Biochem J 340 ( Pt 3):677-86 (1999). 1. Reinhardt, R. L., Hong, S., Kang, S.-J., Wang, Z.-e., and Locksley, R. M. (2006) Visualization of IL-12/23p40 In Vivo Reveals Immunostimulatory Dendritic Cell Migrants that Promote Th1 Differentiation, 177, 1618-1627.

GFP