Recombinant Anti-GFP antibody [E385] (ab32146)
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
Related conjugates and formulations
Overview
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Product name
Anti-GFP antibody [E385]
See all GFP primary antibodies -
Description
Rabbit monoclonal [E385] to GFP -
Host species
Rabbit -
Specificity
This antibody is specific for GFP and GFP fusion proteins. -
Tested applications
Suitable for: WBmore details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Species independent -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Pure GFP protein, or cells known to overexpress GFP.
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General notes
On the basis of low sequence homology, ab32146 is predicted to show no or limited cross-reactivity to RFP and BFP.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E385 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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WB | (3) |
1/1000 - 1/20000.
GFP - 27kDa, Proprietary tag/GFP fusion protein - 52kDa |
Notes |
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WB
1/1000 - 1/20000. GFP - 27kDa, Proprietary tag/GFP fusion protein - 52kDa |
Target
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Relevance
Function: 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. -
Alternative names
- GFP antibody
- Green fluorescent protein antibody
Images
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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
Secondary
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.
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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)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (80)
ab32146 has been referenced in 80 publications.
- Wang Q et al. Rice black-streaked dwarf virus P10 promotes phosphorylation of GAPDH (glyceraldehyde-3-phosphate dehydrogenase) to induce autophagy in Laodelphax striatellus. Autophagy 18:745-764 (2022). PubMed: 34313529
- Lei J et al. Plastidic pyruvate dehydrogenase complex E1 component subunit Alpha1 is involved in galactolipid biosynthesis required for amyloplast development in rice. Plant Biotechnol J 20:437-453 (2022). PubMed: 34655511
- Lee B et al. FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury. J Biol Chem 298:101647 (2022). PubMed: 35101451
- Jian Y et al. Toxicity and action mechanisms of silver nanoparticles against the mycotoxin-producing fungus Fusarium graminearum. J Adv Res 38:1-12 (2022). PubMed: 35572400
- Wang J et al. The Plant Homeodomain Protein Clp1 Regulates Fungal Development, Virulence, and Autophagy Homeostasis in Magnaporthe oryzae. Microbiol Spectr 10:e0102122 (2022). PubMed: 36036638