Mouse Green Fluorescent Protein Monoclonal Antibody | anti-GFP antibody

Green Fluorescent Protein (GFP) (Biotin)

Cat. Num. AAA6121180

• Applications: Immunohistochemistry, Western Blot, ELISA
• Purity: Purified by Protein G Affinity Chromatography
• Synonyms: Green Fluorescent Protein; Monoclonal Antibody; Green Fluorescent Protein (GFP) (Biotin); anti-GFP antibody

• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1
• Clone Number: 168AT1211
• Specificity: Recognizes Green Fluorescent Protein
• Purity/Purification: Purified by Protein G Affinity Chromatography
• Form/Format: Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with Biotin.

• Applicable Applications for anti-GFP antibody: Immunohistochemistry (IHC), Western Blot (WB), ELISA (EIA)
• Application Notes: ELISA: 1:3,000; IHC: 1:50-100; WB: 1:2,000; Applications are based on unconjugated antibody.
• Immunogen: Purified His-tagged GFP protein.
• Conjugate: Biotin
• Preparation and Storage: Store product at 4 degree C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20 degree C. Aliquots are stable at -20 degree C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Western Blot (WB)

(Western blot analysis of anti-GFP Mab 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.)

Western Blot (WB)

(Western blot analysis of anti-GFP Tag Antibody in GFP recombinant protein. GFP recombinant protein (arrow) was detected using the purified Mab.)

Immunofluorescence (IF)

(Immuno-fluorescence for GFP Tag Antibody. 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.)

Immunohistochemistry (IHC)

(Formalin-fixed and paraffin-embedded mouse skin tissue expression GFP reacted with GFP Tag Antibody, 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.)

Related Product Information for anti-GFP antibody

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. Our 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. Our anti-GFP monoclonal antibody can detect both GFP and YFP but not BFP (Blue fluorescent protein) by western blotting.

Product Categories/Family for anti-GFP antibody

Antibodies; Epitope Tag; GFP

References

Ward, W. W., et al.(1980) Photochem. Photobiol. 31:611¬?613. Chalfie, M., et al.(1994) Science 263:802¬?805. Wu C, et al. (1997) Gene 190(1):157-62. Wachter RM, et al. (1998) Structure 6:1267-77.

NCBI and Uniprot Product Information

• NCBI GI #: 1169893
• UniProt Accession #: P42212; Q17104; Q27903; Q93125
• NCBI Official Full Name: Green fluorescent protein
• UniProt Protein Name: Green fluorescent protein
• Protein Family: Green fluorescent protein
• UniProt Gene Name: GFP
• UniProt Entry Name: GFP_AEQVI

Uniprot Description

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.