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GFP (Green Fluorescent Protein) Antibody

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Rabbit polyclonal antibody against highly purified soluble recombinant green fluorescent protein (GFP) from Aequorea victoria. This rabbit polyclonal anti-GFP antibody is reactive against variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP and EGFP.

EU2: Affinity purified rabbit anti-GFP polyclonal antibody
Suggested Unit Size: 25 ug (0.025 mg)
Suggested Unit Price: $295 US

Approximately 5.0 mg IgG / ml serum (assayed by binding to protein A agarose). Concentration may vary between bleeds.

Product Use Yes / No Recommended Use
Western Blot (WB) dilute 1/1000 to 1/2500 in 1X BLOTTO or 3% BSA in PBS 
Immunoprecipitation (IP) 1.0 ul crude serum per 10 cm tissue culture dish. Use 10 uL protein A agarose CL4B (Amersham-Pharmacia) to precipitate immune complex 
IP-Western protocols 1.0 uL crude serum per 10 cm tissue culture dish. Use 10 uL protein A agarose CL4B to precipitate immune complex. Run samples on 12.5% SDS-PAGE, transfer proteins to PVDF membrane, probe membrane with GFP antibody at 1/1000 to 1/2500 dilution, use protein A-HRP conjugate as secondary reagent in ECL or ECL Plus detection kit (Amersham-Pharmacia) as recommended 
Immunohistochemistry & Immunofluorescence Dilute anti-GFP 1/200 to 1/1000 in 4% normal donkey serum (Jackson Immunologics). Incubate for 1 hour at 37oC in humidified atmosphere. Process with secondary antibody as recommended by supplier. See immunofluorescence photograph (above) for typical results.

For additional information please consult the following references:

  1. McCabe, James B. and Berthiaume, Luc G. (1999) “Functional roles of NH2-terminal domains in subcellular localization” Mol. Biol. Cell 10, 3771-3786).
  2. Renata Jasinska, Qiu-Xia Zhang, Carlos Pilquil, Jay Dewald, Deirdre A., Dillon, Luc G. Berthiaume, George M. Carman and David W. Waggoner, David N. Brindley “Lipid phosphate phosphohydrolase-1 degrades exogenous glycerolipid and sphingolipid phosphate esters”(1999) Biochem. J. 340, 677-686.
  3. Zhang, Qiu-Xia, Pilquil, Carlos, Dewald, Jay, Berthiaume, Luc G., and Brindley, David N. Identification of structurally important domains of lipid phosphate phosphatase-1: implications for sites of action” (2000) Biochem. J. 345, 181-184.

Crude antibody (serum) can be stored for at least 24 months at -80oC without apparent loss of activity. This antibody can be frozen and thawed a few times but avoid multiple unnecessary freeze-thaw cycles. Once thawed, the antibody is stable up to two weeks at 4oC.

This polyclonal anti-GFP antibody allows detection of 2 to 5 ng recombinant GFP in ECL or ECL Plus (Amersham-Pharmacia) detection protocols in under one minute of exposition to film (when used with antibody that is kept at 4oC. Each bleed is tested for activity in a Western blotting procedure.
N.B. Typically, exposure times required to detect 2 ng - 5 ng GFP are achieved in as little as one to five seconds when the anti-GFP antibody is prepared fresh from the –20 or –80 oC stocks and used at a 1/2500 dilution using the procedure described in Klapper et al. (1992) Biotechniques, 12, 651-654.

We have over 150 ml of frozen anti-GFP antiserum in the –80oC freezer. –80oC shelf-life so far is over three years without apparent loss of signal in our Western blot assays. Please note that stocks/supply can be increased upon request. Please note that we carry out only a small inventory (about 2-5 mgs) of affinity purified rabbit anti-GFP polyclonal antibody but larger amounts can be prepared up on demand.