mFX75 for Western blotting and 
mFX73 for Immunoprecipitation & Immunostaining are available!!!

Anti GFP, Monoclonal Antibody 

Preparation: Prepared by a hybridoma, which is established by fusing mouse myeloma cells, NSO and spleen cells of BALB/c mice immunized with the GFP's partial peptide coupled with BSA after being immunized with GFP. The hybridoma supernatant cultured by Hollow fiber instruent was purified by Protein A affinity chromatography.
Subclass: mouse IgG_2a·κ
Concentration: shown on each label (approximately 1 mg/mL)
Appearance: D-PBS(-) containing 10 mmol/L Hepes (pH 7.4) and 10 mmol/L Sodium Azide
Working Dilution: 1 : 500 ~ 1 : 1,000

C:  HepG2 cell Lysate (5 x 10⁴ cells)
Lane 1: HepG2 cell Lysate (5 x 10⁴ cells) + rGFP 1 ng
Lane 2: HepG2 cell Lysate (5 x 10⁴ cells) + rGFP 10 ng

HepG2 cell Lysate with 1 ng or 10 ng of recombinant GFP protein was subjected to SDS-PAGE, transferred to PVDF membrane, and exposed to X-ray film.
Exposure time: 3 minutes
Monoclonal antibodies supplied from Wako and manufacturer S and N were used as primary antibodies at final concentration of 1 μg/mL, respectively.

Preparation: Prepared by a hybridoma, which is established by fusing mouse myeloma cells, NSO and spleen cells of BALB/c mice immunized with the GFP's partial peptide coupled with BSA after being immunized with GFP. The hybridoma supernatant cultured by protein-free culture was subjected to ultrafiltration with an ultrafilter 30,000-molecular-weight cutoff.
Subclass: mouse IgG_2a·κ   
Concentration: shown on each label (approximately 1 mg/mL)
Appearance: PBS containing 10 mmol/L Hepes (pH 7.4) and 10 mmol/L Sodium Azide
Working Dilution: Immunoprecipitation   1 ~ 5 μg/μL Sepharose
                               Immunostaining          0.5 ~ 1 μg/mL

SDS-PAGE Silver Staining
Sample: applied GFP (0.5 ug/10 μL) and each eluent (10 μL)
Gel: SuperSep^TM 10-20%, 17 well (Wako Cat. #191-13071)
Detection Reagent: Silver Stain II Kit _Wako (Wako Cat. #291-50301)

Lane 1: applied GFP
Lane W,M,B: eluent from each anti-GFP antibody solid-phased support. (Wako's anti-GFP
antibody (mFX73), manufacturer M's and B's antibodies are used, respectively.

Anti-GFP antibodies supplied from each manufacturer were attached to a solid-phase of protein G Sepharose and binding with GFP were compared using an immunoprecipitation assay. Wako's antibody (mFX73) bound to GFP with higher affi nity than antibodies supplied from manufacturers M and B.

C. elegans strain: unc-18(e81) ; unc-18 :: EGFP, L4-stage larvae
Fixation: o/n fi xation in 4% paraformaldehyde / 1% glutaraldehyde at 4 degrees C
Primary antibody: Anti-GFP antibody (mFX73) 1 μg/mL + Anti-Serotonin serum (1:500)
Secondary antibody: Cy3 conjugated Anti-mouse antibody (1:200) + Biotin-conjugated Anti-rabbit IgG (1:200) / ABC complex (1:100)
Detection: DAB / Fluorescence