Product nameAnti-GFP antibody [6AT316]
See all GFP primary antibodies
DescriptionMouse monoclonal [6AT316] to GFP
SpecificityDetects both GFP and YFP by Western Blotting.
Tested applicationsSuitable for: IHC-Fr, ELISA, WB, IHC-Pmore details
Species reactivityReacts with: Species independent
Full length native protein (purified) corresponding to GFP.
Database link: P42212
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Storage bufferPreservative: 0.09% Sodium azide
Concentration information loading...
PurityProtein G purified
Purification notesThis antibody was purified by protein G and Ni-NTA nickel affinity chromatography, followed by dialysis against PBS.
Our Abpromise guarantee covers the use of ab38689 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/4000. Predicted molecular weight: 27 kDa.|
|IHC-P||1/50 - 1/100.|
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse skin tissue labelling GFP with ab38689. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 38°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A peroxidase-conjugated goat anti-mouse polyclonal (ready to use) was used as the secondary antibody. -ve shows negative staining of the antibody in a wild type mouse, whereas +ve shows positive staining in a mouse expressing GFP under the LGR5 promoter.
Immunohistochemistry (Frozen sections) analysis of E10.5 mouse embryo tissue sections labeling GFP with ab38689 at 1/200 dilution. The tissue was fixed with paraformaldehyde and permeabilized with PBS / 0.5% v/v Triton X-100. The tissue was incubated with ab38689 in 1% BSA + 5% goat serum + PBST for 12 hours at 25°C. A polyclonal goat anti-mouse Alexa Fluor® 568 secondary antibody was used at 1/1000 dilution.
Anti-GFP antibody [6AT316] (ab38689) at 1/4000 dilution + GFP recombinant protein lysate at 35 µg
Peroxidase-conjugated goat anti-mouse IgG (H+L) at 1/5000 dilution
Predicted band size: 27 kDa
Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes : Anti-GFP antibody [6AT316] (ab38689)
Lane 1 : GFP at 1 µg
Lane 2 : GFP at 0.1 µg
Lane 3 : GFP at 0.01 µg
Lane 4 : YFP at 1 µg
Lane 5 : YFP at 0.1 µg
Lane 6 : YFP at 0.01 µg
Predicted band size: 27 kDa
Observed band size: 27 kDa
GFP and YFP were expressed in bacteria.
Formalin-fixed and paraffin-embedded human cancer tissue reacted with ab38689, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining.
ab38689 staining GFP in Chicken spinal cord tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS) for 16 hours at 4°C. A Cy2®-conjugated Donkey anti-mouse monoclonal (1/100) was used as the secondary antibody.
This product has been referenced in:
- Mitra S et al. Dual regulation of lin28a by Myc is necessary during zebrafish retina regeneration. J Cell Biol 218:489-507 (2019). Read more (PubMed: 30606747) »
- Ahmad ST et al. Capicua regulates neural stem cell proliferation and lineage specification through control of Ets factors. Nat Commun 10:2000 (2019). Read more (PubMed: 31043608) »