Product nameAnti-GFP antibody [9F9.F9]
See all GFP primary antibodies
DescriptionMouse monoclonal [9F9.F9] to GFP
SpecificityAssayed by ELISA for direct binding of antigen recognizes wild type and recombinant GFP. Well suited to titrate GFP in solution using either form of the antibody as the capture or detection antibodies. Shows no reactivity against red fluorescence protein (RFP). This antibody is known to cross react with the wild type (wt), recombinant (rGFP) and enhanced (eGFP) forms.
Tested applicationsSuitable for: WB, IHC-Fr, Sandwich ELISA, ICC/IF, IPmore details
Species reactivityReacts with: Species independent
Fusion protein corresponding to GFP aa 1-246. Full Length Fusion Protein. Derived from the jellyfish Aequorea victoria.
Database link: P42212
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.87% Sodium chloride, 0.42% Potassium phosphate
Concentration information loading...
PurityProtein A purified
Purification notesGFP Monoclonal Antibody was prepared from tissue culture supernatant by Protein A affinity chromatography.
Light chain typekappa
sELISA pair antibody
Our Abpromise guarantee covers the use of ab1218 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/3000 - 1/30000.
(See Abreviews). Recognises the immunogen under native conditions and reduced/denatured conditions.
|IHC-Fr||1/1000 - 1/5000.
Works also with eYFP.
|Sandwich ELISA||1/80000 - 1/500000. Can be paired for Sandwich ELISA with Goat polyclonal to GFP (Biotin) (ab6658).
ab1218 can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated monoclonal anti-GFP is well suited to titrate GFP in a sandwich ELISA in combination with polyclonal anti-GFP as a capture antibody.
|ICC/IF||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration. PubMed: 16135826|
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
All lanes : Anti-GFP antibody [9F9.F9] (ab1218) at 1 mg/ml (1 hour at room temperature)
Lane 1 : HeLa lysate 50ng with BLOTTO, overnight at 4°C
Lane 2 : HeLa lysate 100ng with BLOTTO, overnight at 4°C
Lane 3 : HeLa lysate 500ng with BLOTTO, overnight at 4°C
Blocking peptides at 5 % per lane.
All lanes : IRDye® 800 conjugated Goat anti-mouse IgG (H+L), 45 minutes at room temperature at 1/2500 dilution
Additional bands at: 27 kDa. We are unsure as to the identity of these extra bands.
TIRFM responses to prolonged stimulation with 10 mM Ca2+
C. Cells expressing BS-wt were stimulated with 0.5 or 10 mM Ca2+ for 60 min, 37°C, followed by fixation, permeabilization, and staining with anti-GFP antibody (secondary antibody tagged with Alexa-568). Confocal images of SEP and anti-GFP fluorescence are shown along with the merged image. D. Experiment as in C for cells expressing BS-5A mutant.
HEK-293 cells were seeded on glass coverslips and transiently transfected with BS-wt or BS-5A cDNAs. Cells were incubated in DMEM containing 0.5 or 10 mM Ca2+ for 1 hr at 37°C, 5% CO2. Following incubation, cells were immediately fixed in 4% paraformaldehyde for 10 min, RT, permeabilized with 0.2% Triton X-100 (10 min), blocked with 5% serum and 0.1% BSA then incubated with monoclonal anti-GFP antibody (Abcam), followed by goat anti-mouse secondary antibody conjugated to Alexa-568.
Immunoprecipitation using ab1218 at 0.01µl/µl HEK (human embryonic kidney) 293 whole cell lysate incubated with protein G matrix for 1 hour at 4°C.
Total protein input was 3,000,000 cells.
Lane 1: GFP-RhoA wt
Lane 2: GFP-Rac1 wt
Lane 3: GFP-empty
The western blot antibody was ab290 (undiluted).
ab1218 staining GFP in human prostate epithelial primary cells by Immunocytochemistry/ Immunofluorescence. Cells (ARPE-19 cells transfected with mouse Rab27A-GFP) were fixed with paraformaldehyde, permeabilized with 0.5% Triton x100 and blocking with 5% serum was performed for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500: in 1% goat serum, 0.1%TX100, 1XPBS) for 16 hours at 4°C. An Alexa Fluor®546-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/500 dilution. DAPI was used to stain the cell nuclei (blue) and mRab27A-GFP with ab1218 (red).
ab1218 staining GFP in Monkey Kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 3% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/500) for 12 hour at 4°C. An undiluted Alexa Fluor® 488-conjugated Goat polyclonal was used as the secondary antibody
All lanes : Anti-GFP antibody [9F9.F9] (ab1218) at 1/1000 dilution (2 hours at 25°C)
Lane 1 : J774 with Milk, 2 hours at 25°C
Lane 2 : J774+pEGFP-N1 with Milk, 2 hours at 25°C
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
All lanes : Goat anti-mouse HRP at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Additional bands at: 26 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
This product has been referenced in:
- Safiulina D et al. Miro proteins prime mitochondria for Parkin translocation and mitophagy. EMBO J 38:N/A (2019). Read more (PubMed: 30504269) »
- Choi H et al. Amyloid ß-induced elevation of O-GlcNAcylated c-Fos promotes neuronal cell death. Aging Cell 18:e12872 (2019). Read more (PubMed: 30515991) »