Product nameAnti-GFP antibody
See all GFP primary antibodies
DescriptionGoat polyclonal to GFP
SpecificityAnti-GFP assayed by ELISA for direct binding of antigen recognizes wild type, recombinant and enhanced forms of GFP.
Tested applicationsSuitable for: WB, IP, ELISA, ICC/IF, IHC-P, IHC-FrFl, IHC-Frmore details
Species reactivityReacts with: Species independent
Fusion protein corresponding to Aequorea victoria GFP aa 1-246.
Database link: P42212
- IHC: E5.5 Hex-GFP transgenic mouse embryo. WB: HeLa cells. Green Fluorescent protein.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
Purification notesThis product 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.
Our Abpromise guarantee covers the use of ab6673 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/400 - 1/2000.
(for immunoprecipitated GFP, see Abreview).
|IP||Use at an assay dependent concentration.|
This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP.
|IHC-P||1/200 - 1/1000.|
|IHC-FrFl||Use at an assay dependent concentration.|
|IHC-Fr||1/200 - 1/1000.|
RelevanceFunction: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.
Subunit structure: Monomer.
Tissue specificity: Photocytes.
Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.
Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.
Sequence similarities: Belongs to the GFP family.
Biophysicochemical properties: Absorption: Abs(max)=395 nm
Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm.
- GFP antibody
- Green fluorescent protein antibody
Pth4:eGFP transgenic zebrafish embryos at 1 and 2 dpf were fixed with 4% PFA and washed in PBST. They were then washed in PBDT (1% BSA, 1% DMSO, 0.1% Triton X-100 in PBS, pH 7.4), blocked in 10% normal goat serum/PBDT, and incubated overnight at 4°C with primary antibodies to HuC/D (1/100) and GFP (1/400, Abcam ab6673). Further PBST washes and blocking were followed by secondary antibodies overnight at 4°C. Hoechst 34580 was added to stain nuclei (1/2500). After further PBDT and PBS washes, embryos were mounted for confocal imaging.
Abbreviation: e, eye; hy, hypothalamus; m, midbrain; sc, spinal cord. Scale bars: 100 μm (A-C) 50 μm (D-G).
In utero electroporation of Disc1 and Disc1-100P constructs into wild-type neocortex and analysis at P21.
(Panels D-E”) Expression of the constructs was assessed.
(Panels D-D'') 2 days after transfection in vitro.
(Panels E-E'') at P21 in vivo.
Immunochemistry for FLAG and GFP showed that constructs encoding either WT Disc1, the Disc1-100P variant, or GFP alone, expressed these protein species in transfected HEK-293 cells in vitro (Fig 5D–5D”) and in P21 postmitotic cortical neurons in vivo (Fig 5E–5E”)
Immunofluorescence for assessment of GFP+ myofibers in rat tissue.
VML affected muscle from the 50% MG + HA+LMN group were probed for the presence of GFP. GFP+ fibers were detected in a qualitatively similar magnitude at both 2 and 8 weeks post-injury indicating viable engraftment of donor derived muscle progenitor cells. Scale bars are 1mm for whole mount images, 50 μm for regions of interest.
A portion of the TA muscle from the defect region was embedded in a talcum-based gel, frozen in 2-methylbutane, and supercooled in liquid nitrogen. Cryosections (8 μm) were prepared and stained using standard protocols for hematoxylin & eosin.
ab6673 used at a 1/100 dilution.
Mouse small intestines were washed with DPBS and fixed overnight at 4°C in Zinc formalin. Following sectioning and tissue deparaffanization, antigen retrieval was performed with 10mM Tris base (pH 9.0) buffer using a pressure cooker.
For immunohistochemistry, sections were quenched of endogenous peroxidases by 3% H2O2, and sequentially blocked with Avidin D, biotin, and protein blocking reagents. Primary antibody incubation was conducted at 4°C overnight. Secondary biotinylated antibody was added at a dilution of 1/200, and incubated 2 hours at room temperature. Finally, sections were stained according to the ABC peroxidase protocol and counterstained with hematoxylin.
ab6673 used at a 1/200 dilution.
Panel D: Representative anti-eGFP immunofluorescence of macroH2A WT and DKO jejunum counterstained with DAPI (blue).
All lanes : Anti-GFP antibody (ab6673) at 1 µg/ml (o/n at 4degC)
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) lysate at 10 µg
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate at 10 µg
Lane 3 : CHO/K1 lysate at 10 µg
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma cell line) lysate at 10 µg
Lane 5 : A431 (Human epidermoid carcinoma cell line) lysate at 10 µg
Lane 6 : Jurkat (Human T cell leukemia cell line from peripheral blood) lysate at 10 µg
Lane 7 : NIH/3T3 (Mouse embryo fibroblast cell line) lysate
Lane 8 : E-coli HCP control, 50 ng
Lane 9 : FLAG Positive control lysate at 10 µg
Lane 10 : Red fluorescent protein, 50 ng
Lane 11 : Green fluorescent protein, 50 ng
Lane 12 : Glutathinoe-S-Transferase protein, 50 ng
Lane 13 : Maltose Binding protein, 50 ng
All lanes : Peroxidase goat secondary antibody, 60 min at RT at 1/30000 dilution
Blocking Buffer: 1% Casein-TTBS for 30 min at RT.
E5.5 Hex-GFP transgenic mouse embryo stained for GFP using ab6673 at 1/500 dilution. Secondary antibody is a fluorochrome 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.
All lanes : Anti-GFP antibody (ab6673) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells
Lane 2 : Mock transfected HeLa cell lysate
Lysates/proteins at 35 µg per lane.
All lanes : IRDye® 800 conjugated Donkey-a-Goat IgG [H&L] at 1/2500 dilution
Additional bands at: 33 kDa. We are unsure as to the identity of these extra bands.
All lanes : Anti-GFP antibody (ab6673) at 1/1000 dilution
Lane 1 : MRC5VA lung fibroblast whole cell lysate overexpressing EGFP alone
Lanes 2-3 : MRC5VA lung fibroblast whole cell lysate overexpressing an EGFP fusion protein
Lysates/proteins at 15 µg per lane.
All lanes : HRP-conjugated anti-goat polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 27,55 kDa why is the actual band size different from the predicted?
Exposure time: 5 seconds
Immunofluorescence of TGN mouse liver labeling GFP on hepatocytes with ab6673.
Immunohistochemistry of GFP transgenic mouse liver labeling GFP with ab6673.
This product has been referenced in:
- Hess DL et al. Perivascular cell-specific knockout of the stem cell pluripotency gene Oct4 inhibits angiogenesis. Nat Commun 10:967 (2019). Read more (PubMed: 30814500) »
- Gómez-Molina C et al. Small Extracellular Vesicles in Rat Serum Contain Astrocyte-Derived Protein Biomarkers of Repetitive Stress. Int J Neuropsychopharmacol 22:232-246 (2019). Read more (PubMed: 30535257) »