Anti-GFP antibody (ab13970)
Chicken polyclonal GFP antibody. Validated in WB, IHC, ICC/IF. Cited in 1298 publication(s). Independently reviewed in 83 review(s). Immunogen corresponding to recombinant full length protein.
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- References (1706)
- Protocols
Overview
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Product name
Anti-GFP antibody
See all GFP primary antibodies -
Description
Chicken polyclonal to GFP -
Host species
Chicken -
Tested applications
Suitable for: IHC-P, WB, IHC - Wholemount, IHC-FrFl, ICC/IF, IHC-Fr, IHC-FoFrmore details -
Immunogen
Recombinant full length protein corresponding to GFP.
Database link: P42212 -
Positive control
- WB: Transgenic mouse spinal cords expressing GFP. ICC/IF: GFP-transfected NIH/3T3 cells.
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
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
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 |
|---|---|---|
| IHC-P | 1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. The concentrations of fixative for the IHC applications were typically 10% formalin or 2% paraformaldehyde. |
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| WB | 1/5000. | |
| IHC - Wholemount | Use at an assay dependent concentration. | |
| IHC-FrFl | Use at an assay dependent concentration. | |
| ICC/IF | 1/2000. Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). |
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| IHC-Fr | 1/1000. | |
| IHC-FoFr | 1/2000. |
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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab13970)Dow et al PLoS One. 2014 Apr 17;9(4):e95236. doi: 10.1371/journal.pone.0095236. eCollection 2014. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Adenoviral Cre induces mosaic activation of rtTA and GFP induction in CAGs-LSL-rtTA3 and CAGs-LSL-RIK animals.
Immunofluorescent stains for GFP and mKate2 in lung sections of triple transgenic mice (CAGs-LSL-rtTA3 or RIK;TG-Ren.713;LSL-KrasG12D). KrasG12D-induced lung adenomas show strong expression of GFP and mKate2.
Lower panel: Higher magnification of the lesion.
White arrows indicate rare cells that show mKate2, but not GFP expression.
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Western blot - Anti-GFP antibody (ab13970)Image is courtesy of an AbReview submitted by Dr Francois Daubeuf.All lanes : Anti-GFP antibody (ab13970) at 1/200 dilution
Lane 1 : Wild type 'naive' HEK293 whole cell lysates
Lanes 2-4 : GFP transfected HEK293 whole cell lysates
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Donkey anti-chicken polyclonal CY5 conjugate. at 1/1000 dilution
Performed under reducing conditions.
Observed band size: 25 kDa why is the actual band size different from the predicted?
Exposure time: 25 minutesCells were treated by mixing in RIPA buffer and denaturating in Laemmli buffer for 5 minutes at 95°C. The gel was Precast at 4-12%. Blocking was with 0.5% milk at 20°C for 5 minutes.
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Immunohistochemistry (Frozen sections) - Anti-GFP antibody (ab13970)This image is courtesy of an anonymous Abreviewab13970 staining mouse olfactory bulb tissue sections by IHC-Fr.
Sections were PFA fixed, permeabilized in 0.4% Triton-X and blocked with 5% serum for 2 hours at 25°C. The primary antibody was diluted 1/1000 and incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 488 conjugated goat anti-chicken was used as the secondary.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab13970)Image courtesy of an anonymous Abreview.ab13970 staining GFP in murine lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then permeabilized with 0.1% Tween, blocked with 15% serum for 30 minutes at 23°C and then incubated with ab13970 at a 1/500 dilution for 14 hours at 4 °C. The secondary used was an Alexa-Fluor® 488 conjugated goat anti-chicken polyclonal used at a 1/500 dilution.
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Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-GFP antibody (ab13970)Image from Groner AC et al, J Biol Chem. 2012 Jul 20;287(30):25361-9. Epub 2012 May 17, Fig 1. DOI 10.1074/jbc.M112.350884 July 20, 2012 The Journal of Biological Chemistry, 287, 25361 -25369.ab13970 staining GFP in murine olfactory bulb tissue by Immunohistochemistry (PFA perfusion fixed frozen sections) Counterstained with DAPI. -
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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Immunohistochemistry (Frozen sections) - Anti-GFP antibody (ab13970)This image is courtesy of an Abreview submitted by Dr Schwob's Labab13970 staining mouse olfactory epithelium tissue sections by IHC-Fr.
The sample was PFA fixed and blocked with 4% BSA/ 5% NFDM/ 10% NDS for 15 minutes at 20°C. The primary antibody was diluted 1/1000 and incubated with the sample for 1 hour. An FITC conjugated goat anti-chicken was used as the secondary.
This colony is the result of retroviral infection with a control virus. The GFP is under the control of an IRES promoter, so its expression is independent of any other protein. The counterstain is hoescht.
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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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Immunohistochemistry (Frozen sections) - Anti-GFP antibody (ab13970)This image is a courtesy of Ben Devermanab13970 staining GFP in mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections).
Tissue samples were fixed with paraformaldehyde and permeabilized with 0.1% Triton X100 before blocking with 10% serum for 30 minutes at 25°C. The sample was incubated with primary antibody (1/2000) for 16 hours at 25°C in 10% NGS in PBS + 0.1% TX100. An Alexa Fluor®488-conjugated Goat polyclonal to chicken IgG was used as secondary antibody at 1/400 dilution.
In the image, green staining represents GFP expressed in oligodendrocytes, blue is for ToPro3.
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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
This product has been referenced in:
- Handara G et al. The role of agrin, Lrp4 and MuSK during dendritic arborization and synaptogenesis in cultured embryonic CNS neurons. Dev Biol 445:54-67 (2019). Read more (PubMed: 30385274) »
- Al-Moyed H et al. A dual-AAV approach restores fast exocytosis and partially rescues auditory function in deaf otoferlin knock-out mice. EMBO Mol Med 11:N/A (2019). Read more (PubMed: 30509897) »
