Anti-GFP antibody (ab6673)
Key features and details
- Goat polyclonal to GFP
- Suitable for: WB, IP, ELISA, ICC/IF, IHC-P, IHC-FrFl, IHC-Fr
- Reacts with: Species independent
- Isotype: IgG
Overview
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Product name
Anti-GFP antibody
See all GFP primary antibodies -
Description
Goat polyclonal to GFP -
Host species
Goat -
Specificity
Anti-GFP assayed by ELISA for direct binding of antigen recognizes wild type, recombinant and enhanced forms of GFP. -
Tested applications
Suitable for: WB, IP, ELISA, ICC/IF, IHC-P, IHC-FrFl, IHC-Frmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Fusion protein corresponding to Aequorea victoria GFP aa 1-246.
Database link: P42212 -
Positive control
- IHC: E5.5 Hex-GFP transgenic mouse embryo. WB: Pure GFP protein, or cells known to overexpress GFP.
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General notes
This anti-GFP antibody cross reacts with eGFP .
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
Preservative: 0.01% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride -
Concentration information loading... -
Purity
Affinity purified -
Purification notes
This product was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. -
Clonality
Polyclonal -
Isotype
IgG -
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 ab6673 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 | (3) |
1/400 - 1/2000.
(for immunoprecipitated GFP, see Abreview). |
| IP |
Use at an assay dependent concentration.
|
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| ELISA |
1/40000.
This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP. |
|
| ICC/IF | (1) |
1/500.
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| IHC-P | (9) |
1/200 - 1/1000.
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| IHC-FrFl |
Use at an assay dependent concentration.
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| IHC-Fr | (6) |
1/200 - 1/1000.
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| IF |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
1/400 - 1/2000. (for immunoprecipitated GFP, see Abreview). |
|
IP
Use at an assay dependent concentration. |
|
ELISA
1/40000. This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP. |
|
ICC/IF
1/500. |
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IHC-P
1/200 - 1/1000. |
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IHC-FrFl
Use at an assay dependent concentration. |
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IHC-Fr
1/200 - 1/1000. |
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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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Immunohistochemistry - Free Floating - Anti-GFP antibody (ab6673)Suarez-Bregua et al PLoS One. 2017 Oct 17;12(10):e0186444. doi: 10.1371/journal.pone.0186444. eCollection 2017. Fig 1. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Pth4:eGFP transgenic zebrafish embryos at 1 and 2 dpf were fixed with 4% PFA and washed in PBST. They were then washed in PBDT (1% BSA, 1% DMSO, 0.1% Triton X-100 in PBS, pH 7.4), blocked in 10% normal goat serum/PBDT, and incubated overnight at 4°C with primary antibodies to HuC/D (1/100) and GFP (1/400, Abcam ab6673). Further PBST washes and blocking were followed by secondary antibodies overnight at 4°C. Hoechst 34580 was added to stain nuclei (1/2500). After further PBDT and PBS washes, embryos were mounted for confocal imaging.
Abbreviation: e, eye; hy, hypothalamus; m, midbrain; sc, spinal cord. Scale bars: 100 μm (A-C) 50 μm (D-G).
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab6673)Borkowska et al PLoS One. 2016 May 31;11(5):e0156082. doi: 10.1371/journal.pone.0156082. eCollection 2016. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/In utero electroporation of Disc1 and Disc1-100P constructs into wild-type neocortex and analysis at P21.
(Panels D-E”) Expression of the constructs was assessed.
(Panels D-D'') 2 days after transfection in vitro.
(Panels E-E'') at P21 in vivo.
Immunochemistry for FLAG and GFP showed that constructs encoding either WT Disc1, the Disc1-100P variant, or GFP alone, expressed these protein species in transfected HEK-293 cells in vitro (Fig 5D–5D”) and in P21 postmitotic cortical neurons in vivo (Fig 5E–5E”)
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Immunohistochemistry (Frozen sections) - Anti-GFP antibody (ab6673)Goldman et al PLoS One. 2018 Jan 12;13(1):e0191245. doi: 10.1371/journal.pone.0191245. eCollection 2018. Fig 5. Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/Immunofluorescence for assessment of GFP+ myofibers in rat tissue.
VML affected muscle from the 50% MG + HA+LMN group were probed for the presence of GFP. GFP+ fibers were detected in a qualitatively similar magnitude at both 2 and 8 weeks post-injury indicating viable engraftment of donor derived muscle progenitor cells. Scale bars are 1mm for whole mount images, 50 μm for regions of interest.
A portion of the TA muscle from the defect region was embedded in a talcum-based gel, frozen in 2-methylbutane, and supercooled in liquid nitrogen. Cryosections (8 μm) were prepared and stained using standard protocols for hematoxylin & eosin.
ab6673 used at a 1/100 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab6673)Cedeno et al PLoS One. 2017 Sep 21;12(9):e0185196. doi: 10.1371/journal.pone.0185196. eCollection 2017. Fig 3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Mouse small intestines were washed with DPBS and fixed overnight at 4°C in Zinc formalin. Following sectioning and tissue deparaffanization, antigen retrieval was performed with 10mM Tris base (pH 9.0) buffer using a pressure cooker.
For immunohistochemistry, sections were quenched of endogenous peroxidases by 3% H2O2, and sequentially blocked with Avidin D, biotin, and protein blocking reagents. Primary antibody incubation was conducted at 4°C overnight. Secondary biotinylated antibody was added at a dilution of 1/200, and incubated 2 hours at room temperature. Finally, sections were stained according to the ABC peroxidase protocol and counterstained with hematoxylin.
ab6673 used at a 1/200 dilution.
Panel D: Representative anti-eGFP immunofluorescence of macroH2A WT and DKO jejunum counterstained with DAPI (blue).
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Western blot - Anti-GFP antibody (ab6673)All lanes : Anti-GFP antibody (ab6673) at 1 µg/ml (o/n at 4degC)
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) lysate at 10 µg
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate at 10 µg
Lane 3 : CHO/K1 lysate at 10 µg
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma cell line) lysate at 10 µg
Lane 5 : A431 (Human epidermoid carcinoma cell line) lysate at 10 µg
Lane 6 : Jurkat (Human T cell leukemia cell line from peripheral blood) lysate at 10 µg
Lane 7 : NIH/3T3 (Mouse embryo fibroblast cell line) lysate
Lane 8 : E-coli HCP control, 50 ng
Lane 9 : FLAG Positive control lysate at 10 µg
Lane 10 : Red fluorescent protein, 50 ng
Lane 11 : Green fluorescent protein, 50 ng
Lane 12 : Glutathinoe-S-Transferase protein, 50 ng
Lane 13 : Maltose Binding protein, 50 ng
Secondary
All lanes : Peroxidase goat secondary antibody, 60 min at RT at 1/30000 dilutionBlocking Buffer: 1% Casein-TTBS for 30 min at RT.
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Immunofluorescence - Anti-GFP antibody (ab6673)E5.5 Hex-GFP transgenic mouse embryo stained for GFP using ab6673 at 1/500 dilution. Secondary antibody is a fluorochrome conjugated anti-goat IgG secondary antibody at 1/10,000 for 45 min at RT.
Staining: GFP as green fluorescent signal with DAPI blue counterstain.
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Western blot - Anti-GFP antibody (ab6673)All lanes : Anti-GFP antibody (ab6673) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells
Lane 2 : Mock transfected HeLa cell lysate
Lysates/proteins at 35 µg per lane.
Secondary
All lanes : IRDye® 800 conjugated Donkey-a-Goat IgG [H&L] at 1/2500 dilution
Additional bands at: 33 kDa. We are unsure as to the identity of these extra bands. -
Western blot - Anti-GFP antibody (ab6673)This image is courtesy of an anonymous abreview.All lanes : Anti-GFP antibody (ab6673) at 1/1000 dilution
Lane 1 : MRC5VA lung fibroblast whole cell lysate overexpressing EGFP alone
Lanes 2-3 : MRC5VA lung fibroblast whole cell lysate overexpressing an EGFP fusion protein
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : HRP-conjugated anti-goat polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 27,55 kDa why is the actual band size different from the predicted?
Exposure time: 5 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab6673)This image is courtesy of Bart RountreeImmunofluorescence of TGN mouse liver labeling GFP on hepatocytes with ab6673.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab6673)This image is courtesy of Jeff KleinImmunohistochemistry of GFP transgenic mouse liver labeling GFP with ab6673.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (320)
ab6673 has been referenced in 320 publications.
- Xu T et al. Myofibroblast induces hepatocyte-to-ductal metaplasia via laminin-?vß6 integrin in liver fibrosis. Cell Death Dis 11:199 (2020). PubMed: 32251270
- Zieger HL et al. Disease-associated synaptic scaffold protein CNK2 modulates PSD size and influences localisation of the regulatory kinase TNIK. Sci Rep 10:5709 (2020). PubMed: 32235845
- Chen L et al. The role of bone marrow-derived cells in the origin of liver cancer revealed by single-cell sequencing. Cancer Biol Med 17:142-153 (2020). PubMed: 32296582
- Morales Torres C et al. Selective inhibition of cancer cell self-renewal through a Quisinostat-histone H1.0 axis. Nat Commun 11:1792 (2020). PubMed: 32286289
- Klepfish M et al. LOXL2 Inhibition Paves the Way for Macrophage-Mediated Collagen Degradation in Liver Fibrosis. Front Immunol 11:480 (2020). PubMed: 32296422