GFP (AA 246) antibody

Antigen: GFP

Binding Site: AA 246

Protein Type: Recombinant

Clonality: Polyclonal

Host: Goat

Reactivity: Aequorea victoria

Conjugate: Un-conjugated

Application: ELISA, Western Blotting (WB), Immunohistochemistry (IHC)

Catalog no.: ABIN100085

Additional Information

UniProt: P42212

Immunogen: The immunogen is a Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence derived from the jellyfish Aequorea victoria.

Format: Liquid

Isotype: IgG

Description: Green fluorescent protein is a 27 kDa protein produced from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelength of 509nm) when excited by blue light. GFP is an important tool in cell biology research. GFP is widely used enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining.

Characteristics: Concentration Definition: by UV absorbance at 280 nm

Application Details

Application Notes: Anti-GFP is designed to detect GFP and its variants. 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. Biotin conjugated polyclonal anti-GFP used in a sandwich ELISA is well suited to titrate GFP in solution using this antibody in combination with monoclonal anti-GFP (600-301-215) using either form of the antibody as the capture or detection antibody. However, use the monoclonal form only for the detection of wild type or recombinant GFP as this form does not sufficiently detect 'enhanced' GFP. The detection antibody is typically conjugated to biotin and subsequently reacted with streptavidin-HRP (code # S000-03). Fluorochrome conjugated polyclonal anti-GFP can be used to detect GFP by immunofluorescence microscopy in prokaryotic (E.coli) and eukaryotic (CHO cells) expression systems and detects GFP containing inserts. Significant amplification of signal is achieved using fluorochrome conjugated polyclonal anti-GFP relative to the fluorescence of GFP alone. For immunoblotting use either alkaline phosphatase or peroxidase conjugated polyclonal anti-GFP to detect GFP or GFP-containing proteins on western blots. Researchers should determine optimal titers for applications.
ELISA Dilution: 1:40,000
Immunohistochemistry Dilution: 1:200 - 1:1,000
Western Blot Dilution: 1:400 - 1:2,000

Concentration: 1.0 mg/mL

Purification: Sterile filtered

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative: 0.01% (w/v) Sodium azide

Storage: Expiration date is one (1) year from date of opening. None. Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Research Area: Tags/Labels

Restrictions: For Research Use only

Images

Western Blot of Rabbit anti-GFP antibody. Lane 1: HeLa cells. Lane 2: mock transfected HeLa cell lysate. Load: 35 µg per lane. Primary antibody: GFP antibody at 1 µg/ml for 1 h at room temperature. Secondary antibody: IRDye® 800 conjugated Donkey-a-Goat IgG [H&L] MX7 (605-732-125) secondary antibody at 1:2,500 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 27 kDa, 33 kDa for GFP. Other band(s): none.

antibodies-online's polyclonal anti-GFP antibody at a 1:1,000 dilution detects tau-GFP in cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) in Drosophila melanogaster late stage embryonic central nervous system. AlexaFluor 488™ conjugated anti-Goat antibody was used for detection at 1:300. Panel A shows a lateral view (ventral left) and Panels B and C show ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up. Personal Communication, Helmata Mistry, Washington University School of Medicine, St. Louis, MO.

anti-GFP (AA 246) antibody (Image 3)

anti-GFP (AA 246) antibody (Image 4)

Publications

Saxena, Saffery, Wong et al.: "Centromere proteins Cenpa, Cenpb, and Bub3 interact with poly(ADP-ribose) polymerase-1 protein and are poly(ADP-ribosyl)ated." in: The Journal of biological chemistry, Vol. 277, Issue 30, pp. 26921-6, 2002 (PubMed).

Balsam, Wagers, Christensen et al.: "Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium." in: Nature, Vol. 428, Issue 6983, pp. 668-73, 2004 (PubMed).

Kofidis, de Bruin, Yamane et al.: "Stimulation of paracrine pathways with growth factors enhances embryonic stem cell engraftment and host-specific differentiation in the heart after ischemic myocardial injury." in: Circulation, Vol. 111, Issue 19, pp. 2486-93, 2005 (PubMed).

Taura, Miura, Iwaisako et al.: "Hepatocytes do not undergo epithelial-mesenchymal transition in liver fibrosis in mice." in: Hepatology (Baltimore, Md.), Vol. 51, Issue 3, pp. 1027-36, 2010 (PubMed).

La: "A dual promoter lentiviral vector for the in vivo evaluation of gene therapeutic approaches to axon regeneration after spinal cord injury." in: Gene therapy, Vol. 17, Issue 5, pp. 577-91, 2010 (PubMed).

Xu, Leinwand, Dell et al.: "The calmodulin-stimulated adenylate cyclase ADCY8 sets the sensitivity of zebrafish retinal axons to midline repellents and is required for normal midline crossing." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 30, Issue 21, pp. 7423-33, 2010 (PubMed).

Maher-Laporte, Berthiaume, Moreau et al.: "Molecular composition of staufen2-containing ribonucleoproteins in embryonic rat brain." in: PLoS ONE, Vol. 5, Issue 6, pp. e11350, 2010 (PubMed).

Chotard, Skorobogata, Sylvain et al.: "TBC-2 Is Required for Embryonic Yolk Protein Storage and Larval Survival during L1 Diapause in Caenorhabditis elegans." in: PLoS ONE, Vol. 5, Issue 12, pp. e15662, 2011 (PubMed).

Product: Justice, Yuan, Sawchenko et al.: "Type 1 corticotropin-releasing factor receptor expression reported in BAC transgenic mice: implications for reconciling ligand-receptor mismatch in the central corticotropin-releasing factor system." in: The Journal of comparative neurology, Vol. 511, Issue 4, pp. 479-96, 2008 (PubMed).

Puthussery, Gayet-Primo, Taylor: "Localization of the calcium-binding protein secretagogin in cone bipolar cells of the mammalian retina." in: The Journal of comparative neurology, Vol. 518, Issue 4, pp. 513-25, 2009 (PubMed).

Hilgen, von Maltzahn, Willecke et al.: "Subcellular distribution of connexin45 in OFF bipolar cells of the mouse retina." in: The Journal of comparative neurology, Vol. 519, Issue 3, pp. 433-50, 2010 (PubMed).

Brett, Renault, Rafalski et al.: "The microRNA cluster miR-106b~25 regulates adult neural stem/progenitor cell proliferation and neuronal differentiation." in: Aging, Vol. 3, Issue 2, pp. 108-24, 2011 (PubMed). Method employed by authors: Immunohistochemistry (IHC) (1:500, Sample species: Mouse (Murine)).