Green Fluorescent Protein (GFP) Antikörper

Antigen: Green Fluorescent Protein (GFP)

Klonalität: Monoklonal (9F9.F9)

Wirt: Maus

Applikation: Immunhistochemie (IHC), ELISA, Western Blot (WB), Immunpräzipitation (IP)

Produktnummer: ABIN129564

Produktbeschreibung

Swiss-Prot: P42212

Immunogen: The immunogen is a GST- Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246 aa) derived from the jellyfish Aequorea victoria.

Format: Liquid (sterile filtered)

Isotyp: IgG2a  (Passende Sekundärantikörper)

Klon: 9F9.F9

Beschreibung: Anti-GFP (MOUSE) Monoclonal Antibody

Spezifität: Monoclonal anti-GFP was raised against full length recombinant GFP. This antibody recognizes all variants of GFP tested to date by Elisa and Western Blot. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Mouse Serum and purified and partially purified Green Fluorescent Protein (Aequorea victoria). Reactivity is observed against wild-type and recombinant forms of GFP. No reaction is seen against RFP.

Anwendungen

Applikationshinweise: This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated monoclonal anti-GFP used in a sandwich ELISA is well suited to titrate GFP in solution using this antibody in combination with a polyclonal anti-GFP (ABIN116749) using either form of the antibody as the capture or detection antibodies. Only use the monoclonal form for the detection of wild type or recombinant GFP as this form does not sufficiently detect 'enhanced' GFP. Polyclonal anti-GFP detects all variants of GFP tested to date. The detection antibody is typically conjugated to biotin and subsequently reacted with streptavidin conjugated HRP (RA021HRP). While not yet tested, this antibody will likely detect GFP and GFP containing proteins by immunofluorescence microscopy, with properties similar to its companion polyclonal antibody. The use of polyclonal anti-GFP results in significant amplification of signal when fluorochrome conjugated polyclonal anti-GFP is used 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. Recommended dilutions: ELISA 1:200,000. Western blot 1:3,000 - 1:30,000. Immunohistochemistry 1:1,000 - 1:5,000. Successful use in immunoprecipitation was reported by some researchers. As this detection method has not been verified by antibodies-online, the application was deleted from the database. This does not necessarily exclude the use in such procedure. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Konzentration: 2.06 mg/ml (by UV absorbance at 280 nm)

Reinigung: This product was prepared from tissue culture supernatant by Protein A affinity chromatography.

Buffer: 0.02M Potassium Phosphate, 0.15M Sodium Chloride, pH 7.2, Stabilizer: None, 0.01% (w/v) Sodium Azide

Lagerung: 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. Expiration date is one (1) year from date of opening.

Forschungsgebiet: Tags/Labels

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Western blot of GFP recombinant protein detected with this monoclonal anti-GFP antibody. GFP recombinant protein was expressed in HeLa cells, where 50 ng (lane 1), 100 ng (lane 2) and 500 ng (lane 3) of lysate were loaded per lane. anti-GFP detects a 27 kDa band corresponding to the epitope tag GFP. The cell lysates were prepared in a RIPA buffer containing 200 mM NaCl. A 4-12% Bis-Tris gradient gel (Invitrogen) was used for SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking with 5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1.0?µg/ml for 1 h at room temperature followed by washes and reaction with a 1:2,500 dilution of IRDye® 800 conjugated Goat-a-Mouse IgG [H&L] MX10 (610-132-121). IRDye® 800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.