Anti-GFP antibody [9F9.F9] (ab1218)
Key features and details
- Mouse monoclonal [9F9.F9] to GFP
- Suitable for: WB, IHC-Fr, Sandwich ELISA, ICC/IF
- Reacts with: Species independent
- Isotype: IgG1
Overview
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Product name
Anti-GFP antibody [9F9.F9]
See all GFP primary antibodies -
Description
Mouse monoclonal [9F9.F9] to GFP -
Host species
Mouse -
Specificity
This antibody is known to cross react with the wild type (WT), recombinant (rGFP) and enhanced (eGFP) forms.
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Tested applications
Suitable for: WB, IHC-Fr, Sandwich ELISA, ICC/IFmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Recombinant fragment within GFP aa 1 to the C-terminus. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements. Full Length Fusion Protein. Derived from the jellyfish Aequorea victoria.
Database link: P42212 -
Positive control
- Pure GFP protein, or cells known to overexpress GFP.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 0.87% Sodium chloride, 0.42% Potassium phosphate -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
GFP Monoclonal Antibody was prepared from tissue culture supernatant by Protein A affinity chromatography. -
Clonality
Monoclonal -
Clone number
9F9.F9 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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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 ab1218 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 | (23) |
1/3000 - 1/30000.
(See Abreviews). Recognises the immunogen under native conditions and reduced/denatured conditions. |
IHC-Fr | (2) |
1/1000 - 1/5000.
Works also with eYFP. |
Sandwich ELISA |
Use at an assay dependent concentration.
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ICC/IF | (13) |
Use at an assay dependent concentration.
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Notes |
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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
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
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 [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.
Secondary
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. -
Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling GFP with ab1218 at 1/100. Cells were fixed with Formaldehyde and permeabilised with 0.2% Triton100 (permeablized after primary incubation). A good anti-mouse Texas Red® at 1/1000 was used as the secondary antibody.
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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.
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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.
Secondary
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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (395)
ab1218 has been referenced in 395 publications.
- Sastourné-Arrey Q et al. Adipose tissue is a source of regenerative cells that augment the repair of skeletal muscle after injury. Nat Commun 14:80 (2023). PubMed: 36604419
- Ge P et al. M. tuberculosis PknG manipulates host autophagy flux to promote pathogen intracellular survival. Autophagy 18:576-594 (2022). PubMed: 34092182
- Keeble AH et al. DogCatcher allows loop-friendly protein-protein ligation. Cell Chem Biol 29:339-350.e10 (2022). PubMed: 34324879
- Combot Y et al. Seipin localizes at endoplasmic-reticulum-mitochondria contact sites to control mitochondrial calcium import and metabolism in adipocytes. Cell Rep 38:110213 (2022). PubMed: 35021082
- Darade AR et al. Effect of mRNA Delivery Modality and Formulation on Cutaneous mRNA Distribution and Downstream eGFP Expression. Pharmaceutics 14:N/A (2022). PubMed: 35057047