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Products:Tags & Cell Markers >> Fusion / Marker Proteins >> GFP
Overview
Product name
Anti-GFP antibody
See all GFP products (29) ...
Description
Chicken polyclonal to GFP
Specificity
This anti-GFP antibody recognizes the enhanced form of GFP, and all of the fluorescent proteins made by Aequorea victoria, including yellow FP.
Tested applications
IHC-P, WB, IHC-FrFl, ICC/IF, IHC-Fr, IHC-FoFrmore details
Immunogen
Recombinant full length protein.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Thimerosal / Merthiolate
Constituents: 50% Glycerol, PBS
Concentration
Concentration information loading...
Purity
IgY fraction
Purification notes
Serile filtered.
Clonality
Polyclonal
Isotype
IgY
Research areas
Applications
Show applications key
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.
IHC-P: 1/500 - 1/1000.Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: 1/5000
IHC-FrFl: Use at an assay dependent concentration.
ICC/IF: Use at an assay dependent dilution. (Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). The concentrations of fixative for the IHC applications were typically 10% formalin or 2% paraformaldehyde.)
IHC-Fr: 1/1000
IHC-FoFr: 1/2000
Target
Relevance
Green fluorescence protein (GFP) is a 27 kDa protein derived from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelength of 509 nm) when excited by blue light (excitation peak at a wavelength of 395 nm). GFP has become an invaluable tool in cell biology research, since its intrinsic fluorescence can be visualized in living cells. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. Other applications of GFP include assessment of protein protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. GFP technology has considerably contributed to a greater understanding of cellular physiology. YFP differs from GFP due to a mutation at T203Y; antibodies raised against full-length GFP should also detect YFP and other variants.
Alternative names
- GFP antibody
- Green Fluorescent Protein antibody
Anti-GFP antibody images:
Immunocytochemistry/ Immunofluorescence - 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 a FITC-labeled goat anti-chicken
Western blot - GFP antibody (ab13970)
Western 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.
Immunohistochemistry (Frozen sections) - GFP antibody (ab13970)
ab13970 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. A 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 counter-stain is hoescht.
This image is courtesy of an Abreview submitted by Dr Schwob Lab
Immunohistochemistry (Frozen sections) - GFP antibody (ab13970)
ab13970 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.
This image is courtesy of an anonymous Abreview
Immunocytochemistry/ Immunofluorescence - GFP antibody (ab13970)
ab13970 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.
This image is courtesy of an Abreview submitted by Dr Radbod Darabi
Western blot - GFP antibody (ab13970)
All lanes : Anti-GFP antibody (ab13970) at 1/500 dilution
Lane 1 : Mouse mammary carcinoma cell line clone expressing GFP - whole cell lysate
Lane 2 : Mouse mammary carcinoma cell line clone expressing GFP - whole cell lysate
Lane 3 : Mouse mammary carcinoma cell line clone expressing GFP - whole cell lysate
Lane 4 : Mouse mammary carcinoma cell line clone expressing GFP - whole cell lysate
Lane 5 : Mouse mammary carcinoma cell line clone expressing GFP - whole cell lysate
Lane 6 : Parental cell line of mouse mammary carcinoma clones, not expressing GFP - whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
HRP conjugated goat anti-chicken at 1/2000 dilution
developed using the ECL technique
Performed under reducing conditions.
Observed band size : 27-30 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
Results from several clones of mouse mammary carcinoma cell line which express GFP (left) and the parental cell line which does not express GFP (far right). This chicken antibody detects GFP in different forms which may represent species specific modifications (see review with human cells). Beta-actin (~47 kD) is shown as loading control.
This image is courtesy of an anonymous Abreview
Immunohistochemistry (Frozen sections) - GFP antibody (ab13970)
ab13970 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 250C. The sample was incubated with primary antibody (1/2000) for 16 hours at 250C 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.
This image is a courtesy of Ben Deverman
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab13970)
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.
Image courtesy of an anonymous Abreview.
References for Anti-GFP antibody (ab13970)
This product has been referenced in:
- Gallio Met al. The coding of temperature in the Drosophila brain. Cell 144:614-24 (2011). IHC (PFA fixed).Read more (PubMed: 21335241) »
- Kochubey O & Schneggenburger R Synaptotagmin increases the dynamic range of synapses by driving ca(2+)-evoked release and by clamping a near-linear remaining ca(2+) sensor. Neuron 69:736-48 (2011). IHC-FrFl.Read more (PubMed: 21338883) »
See all 46 publications for this product
Publishing research using ab13970? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"