Anti-GFP antibody (ab13970)
Key features and details
- Chicken polyclonal to GFP
- Suitable for: WB, ICC/IF
- Reacts with: Species independent
- Isotype: IgY
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
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
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Recombinant full length protein corresponding to GFP.
Database link: P42212 -
Positive control
- ICC: GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line). WB: Transgenic mouse spinal cords.
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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. 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.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (13) |
1/5000.
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| ICC/IF | (22) |
1/2000.
Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). |
| Notes |
|---|
|
WB
1/5000. |
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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
Datasheets and documents
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SDS download
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Datasheet download
References (2684)
ab13970 has been referenced in 2684 publications.
- Horita N et al. Delta-like 1-Expressing Cells at the Gland Base Promote Proliferation of Gastric Antral Stem Cells in Mouse. Cell Mol Gastroenterol Hepatol 13:275-287 (2022). PubMed: 34438113
- La Rocca G et al. Inducible and reversible inhibition of miRNA-mediated gene repression in vivo. Elife 10:N/A (2021). PubMed: 34463618
- Hardcastle BJ et al. A visual pathway for skylight polarization processing in Drosophila. Elife 10:N/A (2021). PubMed: 33755020
- Feng S et al. Scarless engineering of the Drosophila genome near any site-specific integration site. Genetics 217:N/A (2021). PubMed: 33772309
- Schick E et al. Early cis-regulatory events in the formation of retinal horizontal cells. Dev Biol 476:88-100 (2021). PubMed: 33774011