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Catalog #
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Source:
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Lot #
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Size:
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AM1009a
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Mouse
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SDG070403D
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0.1 mg
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Concentration:
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Accession:
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Clone Name:
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Isotype:
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0.25 mg/ml
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168AT1211
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Mouse IgG1
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Applications:
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Reactivity:
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MW (kDa):
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WB, IF, E
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N/A
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Target/Specificity:
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Purified His-tagged GFP protein was used to produced this monoclonal antibody.
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Other Names:
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Green Fluorescent Protein
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Application Data:
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(Top)Western blot analysis of anti-GFP Mab (Cat. #AM1009a) using purified GFP, YFP and BFP proteins expressed in bacteria: Both GFP (Lanes 1-3) and YFP (Lanes 4-6) but not BFP (data not shown) were detected using the purified Mab.
(BUttom)Western blot analysis of anti-GFP Tag Antibody (SG071130A) in GFP recombinant protein. GFP recombinant protein (arrow) was detected using the purified Mab. |
Immuno-fluorescence for GFP Tag Antibody (Cat # AM1009A). Antibody target is a YFP tagged protein (the inner membrane complex) of Toxoplasma gondii, which localizes around the exterior of the parasite. Brighter parasites represent actively dividing cells and darker ones are not actively dividing.
Secondary goat-anti-mouse and goat-anti-rabbit were conjugated with Alexa546 (Molecular Probes)at 1:200 dilution. Phase contrast enhancement applied. Fluorescent lamp exposure times as follows: RFP Channel, 0.075 sec; YFP Channel, 1.000 sec; DAPI Channel, 0.020 sec; Phase Channel: 0.300 sec. Microscope: Axiovert 200M inverted microscope (Zeiss) equipped with a PRIOR stage and a NEOFLUAR 100x/1.3 objective lens. Images were collected with a Hamamatsu C4742-95 CCD camera. Chroma Filtersets: TRITC, HQ545/30x Q570LP HQ620/60m; DAPI, D350/50x E420LP 400DCLP; YFP, Q151LP HQ500/20x HQ535/30m. Data courtesy of Seth Robertson, Boston College. |
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Background:
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Green fluorescent protein (GFP), originally isolated from the jellyfish Aequorea victoria, is one of the best visual reporters for monitoring gene expression in vivo and in situ. GFP is a also convenient marker for use in flow cytometry because it eliminates the need to incubate with a
secondary reagent (such as dyes or antibodies) for detection. However, anti-GFP antibody is also widely used for co-immunipreciapitation, co-localization or western blotting for the confirmation of specificity when a GFP fusion protein is expressed in cells. Abgent's anti-GFP monoclonal antibody provides a simple solution to detect the expression of a GFP-tagged protein in cells. Because of its ability to spontaneously generate its own fluorophore, the green fluorescent protein (GFP) from the jellyfish Aequorea victoria is used extensively as a fluorescent marker in molecular and cell biology. The yellow fluorescent proteins (YFPs) have the longest wavelength emissions of all GFP variants examined to date. This shift in the spectrum is the result of a T203Y substitution (single-letter amino acid code), a mutation rationally designed on the basis of the X-ray structure of GFP S65T. Abgent's anti-GFP monoclonal antibody can detect both GFP and YFP but not BFP (Blue fluorescent protein) by western blotting.
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Background References:
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- Ward, W. W., et al.(1980) Photochem. Photobiol. 31:611613.
- Chalfie, M., et al.(1994) Science 263:802805.
- Wu C, et al. (1997) Gene 190(1):157-62.
- Wachter RM, et al. (1998) Structure 6:1267-77.
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Product Citations:
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- Jinhyun Kim, et al. J Neurophysiol, Oct 2008; 100: 1835 - 1847.
- Witola, William H., et al. J. Biol. Chem. 2006 Jul 28;281(30):21305-21311. (Applications: IF)
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Application Notes:
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| ELISA | 1:3,000 |
| Western blotting | 1:2,000 |
| Immunohistochemistry | 1:50~100 |
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Format:
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Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified by protein G and Ni-NTA nickel affinity chromatography, followed by dialysis against PBS.
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Storage:
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Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.
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Precautions:
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This product is for research use only. Not for use in diagnostic
or therapeutic procedures.
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