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Recombinant full length protein corresponding to GFP aa 1-246.
MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTL VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYN SHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLAD HYQQNTPIGDGPVL LPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK
Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation.
Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC) Biotin/Protein Ratio: 10-20 BAC molecules per goat IgG molecule.
Our Abpromise guarantee covers the use of ab6658 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000 - 1/10000.|
|IP||Use at an assay dependent concentration.|
|ICC/IF||1/1000 - 1/5000.|
|Sandwich ELISA||1/10000 - 1/50000. Can be paired for Sandwich ELISA with Mouse monoclonal [9F9.F9] to GFP (ab1218).
May be used as capture or detection antibody in a Sandwich ELISA.
|In situ hybridization||Use at an assay dependent concentration. PubMed: 24262147|
ab6658 staining GFP in human melanoma cells recovered from nude mice by Immunocytochemistry/ immunoflurescence. Cells were fixed with formaldehyde, permeabilized with 0.25% Triton X-100 RT for 10min and blocking with commercially available blocking buffer was performed for 30 minutes at RT. Samples were incubated with primary antibody (1:50) for 18 hours at 4°C. An Alexa Fluor®488-conjugated donkey polyclonal to goat IgG was used as secondary antibody at 1/100 dilution. Green color indicates GFP/Fibrolast cells, while red color indicates Ki67 positive cells, most of them are tumor cells (Abcam`s ab15580 was used for the detection).
Immunofluorescence Microscopy using ab6658. Tissue: Drosophila melanogaster late stage embryonic central nervous system. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: Anti-GFP antibody at a 1/1,000 for 1 h at RT. Secondary antibody: AlexaFluor 488™ conjugated anti-Goat antibody at 1/300 for 45 min at RT. Panel A: shows a lateral view (ventral left). Panels B and C: shows ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up. Staining: tau-GFP cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) as green fluorescent signal.
Immunofluorescence Microscopy using ab6658. Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Primary antibody: Goat anti- GFP was used at 1/500 dilution in free floating imunnohistochemistry to detect GFP. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:10,000 for 45 min at RT.Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.
ab6658 staining GFP in rat brain cells infected with viruses containing GFP under a CMV promoter by Immunocytochemistry/ Immunofluorescence.Cells were fixed with formaldehyde, permeabilized using 0.2% Triton, blocked with 20% serum and then incubated with ab6658 at a 1:50 dilution for 20 hours at 25°C. The secondary used was an Alexa-Fluor 488 conjugated rabbit polyclonal, used at a 1/200 dilution.
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