Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E385] to GFP
- Suitable for: WB
- Reacts with: Species independent
Product nameAnti-GFP antibody [E385]
See all GFP primary antibodies
DescriptionRabbit monoclonal [E385] to GFP
SpecificityThis antibody is specific for GFP and GFP fusion proteins.
Tested applicationsSuitable for: WBmore details
Unsuitable for: ICC/IF or IP
Species reactivityReacts with: Species independent
within Aequorea victoria GFP aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P42212
- HeLa whole cell lysate (ab29545) can be used as a positive control in WB.
On the basis of low sequence homology, ab32146 is predicted to show no or limited cross-reactivity to RFP, YFP, BFP, and CFP.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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.
Dissociation constant (KD)KD = 1.02 x 10 -12 M Learn more about KD
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab32146 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/20000.
GFP - 27kDa, Proprietary tag/GFP fusion protein - 52kDa
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 [E385] (ab32146) at 1/20000 dilution (purified)
Lane 1 : HeLa whole cell lysate (negative control)
Lane 2 : HeLa whole cell lysate spike with recombinant proprietary tag-GFP fusion protein at 0.001 µg
Lane 3 : HeLa whole cell lysate spike with recombinant proprietary tag-GFP fusion protein at 0.002 µg
Lane 4 : HeLa whole cell lysate spike with recombinant proprietary tag-GFP fusion protein at 0.0025 µg
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 27 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM /TBST.
All lanes : Anti-GFP antibody [E385] (ab32146) at 1/20000 dilution (unpurified)
Lane 1 : HeLa cells lysate (negative control)
Lane 2 : HeLa cell lysate spike with recombinant Proprietary tag-GFP fusion protein at 0.1ng
Lane 3 : HeLa cell lysate spike with recombinant Proprietary tag-GFP fusion protein at 0.5ng
Lane 4 : HeLa cell lysate spike with recombinant Proprietary tag-GFP fusion protein at 1ng
Lane 5 : HeLa cell lysate spike with recombinant Proprietary tag-GFP fusion protein at 2ng
Lane 6 : HeLa cell lysate spike with recombinant Proprietary tag-GFP fusion protein at 2.5ng
Lane 7 : Proprietary tag-GFP 0.1mg/ml (positive control)
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32146 has been referenced in 43 publications.
- Liu N et al. Involvement of the cysteine protease BcAtg4 in development and virulence of Botrytis cinerea. Curr Genet 65:293-300 (2019). PubMed: 30167777
- Liu Z et al. A phosphorylated transcription factor regulates sterol biosynthesis in Fusarium graminearum. Nat Commun 10:1228 (2019). PubMed: 30874562
- Liu N et al. The b-ZIP transcription factor FgTfmI is required for the fungicide phenamacril tolerance and pathogenecity in Fusarium graminearum. Pest Manag Sci N/A:N/A (2019). PubMed: 31025482
- Chandler LC et al. Enhancement of Adeno-Associated Virus-Mediated Gene Therapy Using Hydroxychloroquine in Murine and Human Tissues. Mol Ther Methods Clin Dev 14:77-89 (2019). PubMed: 31309129
- Youseff BH et al. TRAF6 Plays a Proviral Role in Tick-Borne Flavivirus Infection through Interaction with the NS3 Protease. iScience 15:489-501 (2019). PubMed: 31129244