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GFP Tag AntibodyPurified Mouse Monoclonal Antibody (Mab)
| Country | United States
Ordering Information
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| Catalog # | Size | Availability | Price | |
| AM1009a | 0.1 mg 400 ul | In Stock | $ 195.00 | DISCONTINED INQUIRE CLICK INQUIRE Add to cart |
- Specification
- Citiations : 3
- Reviews
- Protocols
- Backgrounds
GFP Tag Antibody - Product info | |
| Application | WB, IF, IHC
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| Concentration | 0.25 mg/ml |
| Isotype | Mouse IgG1 |
| Clone Names | 168AT1211 |
| Calculated MW | 48 kDa |
GFP Tag Antibody - Additional info | |
| Target/Specificity Purified His-tagged GFP protein was used to produced this monoclonal antibody. | |
| Dilution WB~~1:2,000 IF~~1:50~100 IHC~~1:50~100 | |
| Format 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. | |
| Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. | |
| Precautions GFP Tag Antibody is for research use only and not for use in diagnostic or therapeutic procedures. | |
GFP Tag Antibody - Application data
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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.
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Western blot analysis of anti-GFP Tag Antibody (Cat. #AM1009a) in GFP recombinant protein. GFP recombinant protein (arrow) was detected using the purified Mab.
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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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Formalin-fixed and paraffin-embedded mouse skin tissue expression GFP reacted with GFP Tag Antibody (Cat.#AM1009a), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. -ve shows negative staining of the antibody in a wild type mouse, whereas +ve shows positive staining in a mouse expressing GFP under the LGR5 promoter. The staining shows localization of GFP at the bottom of the hair follicle. The antigen was retrieved in citrate pH 6 in water bath for 25 mins.
GFP Tag Antibody - Research Areas
- Witola WH et al. Localization of the Phosphoethanolamine Methyltransferase of the Human Malaria Parasite Plasmodium falciparum to the Golgi Apparatus.J Biol Chem. 2006 Jul 28;281(30):21305-11. Epub 2006 May 16.
- Rooj AK et al. Glioma-specific Cation Conductance Regulates Migration and Cell Cycle Progression. J Biol Chem. 2012 Feb 3;287(6):4053-65.
- Yalcin A et al. Nuclear Targeting of 6-Phosphofructo-2-kinase (PFKFB3) Increases Proliferation via Cyclin-dependent Kinases.J Biol Chem. 2009 Sep 4;284(36):24223-32. doi: 10.1074/jbc.M109.016816.
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Provided below are standard protocols that you may find useful for product applications.
BACKGROUND
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.
REFERENCES
Ward, W. W., et al.(1980) Photochem. Photobiol. 31:611
