Anti-GFP antibody (ab13970)
Key features and details
- Chicken polyclonal to GFP
- Suitable for: WB, ICC/IF
- Reacts with: Recombinant fragment
- Isotype: IgY
Overview
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Product name
Anti-GFP antibody
See all GFP primary antibodies -
Description
Chicken polyclonal to GFP -
Host species
Chicken -
Specificity
Our GFP antibody does cross-react with the many fluorescent proteins that are derived from the jellyfish Aequorea victoria. These are all proteins that differ from the original GFP by just a few point mutations (EGFP, YFP, mVenus, CFP, BFP etc.).
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Tested Applications & Species
See all applications and species dataApplication Species ICC/IF MouseWB Mouse
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Immunogen
Recombinant full length protein corresponding to GFP.
Database link: P42212
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. -
Storage buffer
pH: 7
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate -
Concentration information loading... -
Purity
IgY fraction -
Purification notes
Sterile filtered. -
Clonality
Polyclonal -
Isotype
IgY -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab13970 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC/IF |
Mouse
|
| WB |
Mouse
|
| Application | Abreviews | Notes |
|---|---|---|
| WB | (13) |
1/5000.
|
| ICC/IF | (22) |
1/2000.
Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). |
| Notes |
|---|
|
WB
1/5000. |
|
ICC/IF
1/2000. Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). |
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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Western blot - Anti-GFP antibody (ab13970)This image is courtesy of an anonymous AbreviewAll lanes : Anti-GFP antibody (ab13970) at 1/2000 dilution (Diluent 1x TBS /4 hours at 4°C)
Lane 1 : 3 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
Lane 2 : 2 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
Lane 3 : 1 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
Lysates/proteins at 25 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (ab150176) at 1/5000 dilution
Performed under reducing conditions.
Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 secondsGel Running Conditions: Reduced Denaturing (15% PAGE)
Detection method: Fluorescent Secondary Antibodies
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)This image is courtesy of an Abreview submitted by Christophe Lachaudab13970 staining GFP in U-2 OS (Human bone osteosarcoma epithelial cell line) cells by ICC/IF.
Cells were paraformaldehyde fixed, permeabilized with 0.5% triton and blocked with 2% antibody dilution buffer for 2 hours. Cells were incubated with the primary antibody (1/1000) for 1 hour at 25°C. An undiluted Alexa Fluor® 488 conjugated Goat anti-chicken polyclonal was used as the secondary antibody.
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Western blot - Anti-GFP antibody (ab13970)Image courtesy of an anonymous Abreview.All lanes : Anti-GFP antibody (ab13970) at 1/1000 dilution
Lane 1 : Whole cell lysate prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
Lane 2 : Whole cell lysate prepared from HeLa cells.
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : IRDye 800CW conjugated goat anti-chicken polyclonal at 1/15000 dilution
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)Transgenic mice expressing GFP selectively in lamina II of the spinal cord.
In the right panels, note the correspondance between the green (rabbit anti-GFP) and red signals (chicken anti-GFP from Abcam) indicating that these two antibody preparations recognized the same gene product. The secondary antibody used with ab13970 was an FITC-labeled goat anti-chicken
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)ab13970 staining GFP in GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line) cells.
The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab13970 at 1/2000 dilution overnight at +4°C followed by incubation with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173), for 1 hour, at 1 μg/ml.
Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH/3T3 cells, the cells upon which ab13970 was applied gave a stronger signal in the 488 channel, indicating that ab13970 is binding to GFP and therefore eliciting signal amplification.
ab13970 was also applied to non-GFP-transfected NIH/3T3 cells, which produced no positive staining, indicating specificity for GFP.Nuclear DNA was labeled with 1.43 μM DAPI (blue).
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Western blot - Anti-GFP antibody (ab13970)Lane 1 : Rabbit anti-GFP
Lane 2 : Anti-GFP antibody (ab13970)
All lanes : Transgenic mouse spinal cordsWestern blot of transgenic mouse spinal cords showing that the rabbit anti-GFP (lane 1) and the chicken anti-GFP (Abcam; lane 2) recognize a band at the same molecular weight.
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)This image is courtesy of an Abreview submitted by Dr Radbod Darabiab13970 staining GFP + tumor in mouse muscle cells by ICC/IF.
Cells were formaldehyde fixed and blocked with 3% BSA for 1 hour at 24°C prior to incubation with the primary antibody (1/500) for 1 hour at 24°C. An Alexa Fluor® 488 conjugated goat anti-chicken was used as the secondary.
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab13970)Image courtesy of an AbReview submitted by Dr Francois Daubeuf.Immunocytochemical/ Immunoflurescence analysis of cytospined HEK-293 cells (Human epithelial cell line from embryonic kidney) transfected with GFP, labeling GFP with ab13970 at 1/200 incubated for 16 hours at 4°C with 1% BSA in PBS. Secondary used was a donkey anti-chicken polyclonal DyLight® 594 at 1/500.
GFP is shown in red (DyLight® 594).
Nuclei are counterstained in blue (DAPI).
The left panel shows HEK-293 cells transfected with GFP and the right panel shows non-transfected HEK-293 cells.
Protocols
References (2036)
ab13970 has been referenced in 2036 publications.
- Krawchuk MB et al. Optogenetic assessment of VIP, PV, SOM and NOS inhibitory neuron activity and cerebral blood flow regulation in mouse somato-sensory cortex. J Cereb Blood Flow Metab 40:1427-1440 (2020). PubMed: 31418628
- Schürmann B et al. A novel role for the late-onset Alzheimer's disease (LOAD)-associated protein Bin1 in regulating postsynaptic trafficking and glutamatergic signaling. Mol Psychiatry 25:2000-2016 (2020). PubMed: 30967682
- Gregson CL et al. A Rare Mutation in SMAD9 Associated With High Bone Mass Identifies the SMAD-Dependent BMP Signaling Pathway as a Potential Anabolic Target for Osteoporosis. J Bone Miner Res 35:92-105 (2020). PubMed: 31525280
- Estrada SM et al. The orphan nuclear receptor Nr4a1 mediates perinatal neuroinflammation in a murine model of preterm labor. Cell Death Dis 11:11 (2020). PubMed: 31907354
- Li C et al. Comprehensive transcriptome analysis of cochlear spiral ganglion neurons at multiple ages. Elife 9:N/A (2020). PubMed: 31913118
