GFP Antibody [FITC]

Western Blot of GFP antibody [FITC]. Lane 1: GFP. Lane 2: None. Load: 50 ng per lane.Primary antibody: None.Secondary antibody: Fluorescein goat secondary antibody at 1:1,000 for 60 min at RT.Block for 30 min at RT.Predicted/Observed size: 28 kDa, 33 kDa for GFP. Other band(s): None.

Tissue: Drosophila melanogaster late stage embryonic central nervous system. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: anti-GFP antibody at a 1:1,000 for 1 h at RT. Secondary antibody: AlexaFluor 488 conjugated anti-Goat antibody at 1:300 for 45 min at RT. Panel A: shows a lateral view (ventral left). Panels B and C: shows ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up. Staining: tau-GFP cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) as green fluorescent signal.

Lane 1: HeLa cells. Lane 2: mock transfected HeLa cell lysate. Load: 35 ug per lane. Primary antibody: GFP antibody at 1 ug/ml for 1 h at room temperature. Secondary antibody: IRDye 800 conjugated Donkey-a-Goat IgG [H&L] MX7 secondary antibody at 1:2,500 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 27 kDa, 33 kDa for GFP. Other band(s): none.

Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Primary antibody: Goat anti-GFP was used at 1:500 dilution in free floating immunohistochemistry to detect GFP. Secondary antibody: Fluorochrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:500 at 1:10,000 for 45 min at RT. Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.

Dot Blot Analysis Using GFP Antibody

• Reactivity: All-NA
• Applications: WB, DB, ELISA, Flow, Func, ICC/IF, IHC, KD, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: FITC

GFP Antibody [FITC] Summary

• Description: For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room
  
  GFP Fluorescein Conjugated Antibody was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, anti-Fluorescein and purified and partially purified Green Fluorescent Protein (Aequorea victoria)
• Immunogen: The immunogen is a Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246aa) derived from the jellyfish Aequorea victoria. (Uniprot: P42212)
• Specificity: No reaction was observed against Human, Mouse and Rat Serum Proteins.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • Flow Cytometry 1:10-1:1000
  • Fluorophore-linked immunosorbent assay >1:20000
  • Immunocytochemistry/ Immunofluorescence 1:500-1:2500
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Knockdown Validated
  • Knockout Validated
  • Proximity Ligation Assay
  • Western Blot >1:10000
• Application Notes: Suitable for immunomicroscopy and flow cytometry or FACS analysis as well as other antibody based fluorescent assays requiring lot-to-lot consistency.

Reactivity Notes

No reaction was observed against Human, or Rat serum proteins. Known Cross Reactivity: rGFP. YFP differs from GFP due to a mutation at Thr203Tyr; antibodies raised against full-length GFP should also detect YFP and other variants. Reactivity in transgenic mice with GFP. Reactivity in human cell lines transfected with a GFP construct.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Sodium Azide
• Purity: Immunogen affinity purified

Alternate Names for GFP Antibody [FITC]

• eGFP
• Enhanced Green Fluorescent Protein
• GFP
• GFPuv
• green fluorescent protein (gfp)
• Green Fluorescent Protein

Background

Green fluorescent protein (GFP) is a 27 kDa protein derived from the bioluminescent jellyfish Aquorea Victoria. GFP can be excited by light in the blue to ultraviolet spectrum and has an excitation peak at 395 nm with an emission peak at 509 nm in the green spectrum. GFP autocatalytically forms a fluorescent pigment from natural amino acids present in the nascent protein. GFP is widely used as a reporter (tag) for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. GFP fluorescence is stable under fixation conditions and antibodies are suitable for a variety of applications including ELISA, ICC/IF, IHC, IP and WB (1-3).

References
1. Shi, C., Pan, F. C., Kim, J. N., Washington, M. K., Padmanabhan, C., Meyer, C. T., . . . Means, A. L. (2019). Differential Cell Susceptibilities to Kras(G12D) in the Setting of Obstructive Chronic Pancreatitis. Cell Mol Gastroenterol Hepatol. doi:10.1016/j.jcmgh.2019.07.001

2. Zhao, S., Fortier, T. M., & Baehrecke, E. H. (2018). Autophagy Promotes Tumor-like Stem Cell Niche Occupancy. Curr Biol, 28(19), 3056-3064.e3053. doi:10.1016/j.cub.2018.07.075

3. Zusso, M., Lunardi, V., Franceschini, D., Pagetta, A., Lo, R., Stifani, S., . . . Moro, S. (2019). Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway. J Neuroinflammation, 16(1), 148. doi:10.1186/s12974-019-1538-9

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for GFP Antibody (NB100-1771)(12)

Publications using NB100-1771

• Dontireddy DA. Identification of pulmonary PDGFRalpha-positive fibroblast specific miRNA and mRNA expression profiles during postnatal lung development Thesis 2015-01-01 (ICC/IF, Mouse)
• Choi I; Wang M; Yoo S et al. Autophagy enables microglia to engage amyloid plaques and prevents microglial senescence Nature cell biology 2023-05-25 [PMID: 37231161]
• Fakhiri J, Schneider MA, Puschhof J et al. Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses Mol Ther Methods Clin Dev 2019-02-16 [PMID: 30766894] (FLOW)
• Chaudhuri D, Artiga DJ, Abiria SA et al. Mitochondrial calcium uniporter regulator 1 (MCUR1) regulates the calcium threshold for the mitochondrial permeability transition Proc Natl Acad Sci U S A 2016-03-16 [PMID: 26976564] (IF/IHC)
• Doerner JF, Delling M, Clapham DE. Ion channels and calcium signaling in motile cilia Elife 2015-12-10 [PMID: 26650848] (IHC-Fr)
• Ntokou A, Klein F, Dontireddy D et al. Characterization of the platelet-derived growth factor receptor-alpha-positive cell lineage during murine late lung development. Am J Physiol Lung Cell Mol Physiol 2015-11-01 [PMID: 26320158] (IHC-Fr)
• ElAli A, Menet R, Bernard M et al. Myeloid dickkopf-1 fuels neurovascular and neuroimmune alterations in ischemic stroke Research Square 2023-08-14 (Immunohistochemistry Free-Floating)
• Hauser J, Grundstrom C, Grundstrom T. Allelic Exclusion of IgH through Inhibition of E2A in a VDJ Recombination Complex J Immunol 2014-02-24 [PMID: 24470503] (PLA)
  
  Details:
  This citation used the FITC version of this antibody.
• Freeman K, Tao W, Sun H et al. In situ three-dimensional reconstruction of mouse heart sympathetic innervation by two-photon excitation fluorescence imaging J Neurosci Methods 2013-09-19 [PMID: 24056230] (IHC-Fr, ICC/IF)
  
  Details:
  This citation used the FITC version of this antibody.
• Lampron A, Pimentel-Coelho PM, Rivest S. Migration of bone marrow-derived cells into the CNS in models of neurodegeneration J Comp Neurol 2013-05-16 [PMID: 23682015] (FLOW)
  
  Details:
  This citation used the FITC version of this antibody.
• Hauser J, Sveshnikova N, Wallenius A et al. B-cell receptor activation inhibits AID expression through calmodulin inhibition of E-proteins PNAS 2008-01-01 [PMID: 18203819]
  
  Details:
  This citation used the FITC version of this antibody.
• Yee, Min et al. Type II epithelial cells are critical target for hyperoxia-mediated impairment of postnatal lung development. Am J Physiol Lung Cell Mol Physiol 291: L1101-L1111. 2006-01-01 [PMID: 16861382]

FAQs for GFP Antibody (NB100-1771). (Showing 1 - 1 of 1 FAQs).

1. NB100-1771 reactivity indicates Mu and NA. Does it react with mice?  ..show answer..
   • This product reacts to GFP. It has been used in transgenic mice, but does not react to mice https://www.ncbi.nlm.nih.gov/pubmed/26650848