Supplied as an aliquot of serum with 0.05% Sodium Azide and 1.25% Sodium Chloride.
Storage
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid freeze / thaw cycles.
General Notes
The total IgG concentration has been determined to be 5 mg/ml, however, as this antibody is provided as whole antiserum, the specific IgG concentration is unknown.
Immunofluorescence analysis of 4% paraformaldehyde fixed HEK293 cells transfected with GFP expressing plasmid, permeabilized with 0.15% Triton, using Anti-GFP Antibody (A290) at a 1:200 dilution (1 hour incubation), followed by Goat Anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor 568) (red) at a 1:1,000 dilution. The nuclear stain is DAPI (blue).
Western blot analysis of HEK293 cell lysates (30µg protein in RIPA buffer) using Anti-GFP Antibody (A290) at a 1:5,000 dilution (1 hour incubation). The lanes contain: (A) HEK293 cells transfected with GFP expressing plasmid, (B) non-transfected HEK293 cells. Detected by chemiluminescence.
Lane 1 & 3: IP from COS-7 cells transfected with EGFP.N-Ras (48 kDa). Lane 2 & 4: IP from untransfected COS 7 cells. Lane 1 & 2: IP using 15µl of Anti-GFP Antibody (A290) conjugated to sepharose beads (0.5 mg IgG per ml of beads). Lane 3 & 4: IP using 15 µl of an unpublished goat polyclonal antibody to GFP conjugated to sepharose beads (1 mg IgG per ml of beads). Immunoprecipitation: COS-7 cell lysates containing 100µg of total protein in 200µl of 0.1% SDS-RIPA buffer with addition of complete protease inhibitor were used for each immunoprecipitation. Cell lysates were incubated with Anti-GFP sepharose beads for 2 hours at 4°C with rocking. Beads were washed. Proteins were eluted with 1% SDS 50 mM HEPES (pH 7.4) at 80°C (15 min). Half of the IP sample was loaded on each lane of 10 % SDS PAGE gel and gel was processed for Western blotting/ECL. Western Blot: Primary Antibody: Anti-GFP antibody [LGB-1] (ab291) at 0.2µg/ml in 5% non-fat milk/TBS-T (1 hour incubation at room temperature). Secondary Antibody: Sheep Anti-Mouse IgG Antibody (HRP) at a 1:20,000 dilution in 5% non-fat milk/TBS-T (1 hour incubation at room temperature). Development with ECL plus (GE Healthcare). Exposure time: 5 seconds.
Immunofluorescent side-by-side comparison between Anti-GFP Antibody (A290) from Antibodies.com and Anti-GFP Antibody (ab290) from Abcam. Immunofluorescence analysis of 4% paraformaldehyde fixed HEK293 cells transfected with GFP expressing plasmid, permeabilized with 0.15% Triton, using Anti-GFP Antibody (A290) (Left Panel) and Anti-GFP Antibody (ab290) (Right Panel) at a 1:200 dilution (1 hour incubation), followed by Goat Anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor 568) (red) at a 1:1,000 dilution. The nuclear stain is DAPI (blue). Discover more, for less.
Immunofluorescent side-by-side comparison between Anti-GFP Antibody (A290) from Antibodies.com and Anti-GFP Antibody (A11122) from Thermo Fisher. Immunofluorescence analysis of 4% paraformaldehyde fixed HEK293 cells transfected with GFP expressing plasmid, permeabilized with 0.15% Triton, using Anti-GFP Antibody (A290) (Left Panel) and Anti-GFP Antibody (A11122) (Right Panel) at a 1:200 dilution (1 hour incubation), followed by Goat Anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor 568) (red) at a 1:1,000 dilution. The nuclear stain is DAPI (blue). Discover more, for less.
Western blot side-by-side comparison between Anti-GFP Antibody (A290) from Antibodies.com, Anti-GFP Antibody (ab290) from Abcam, and Anti-GFP Antibody (A11122) from Thermo Fisher. Western blot analysis of HEK293 cell lysates (30µg protein in RIPA buffer) using Anti-GFP Antibody (A290) (Right Panel), Anti-GFP Antibody (ab290) (Middle Panel), and Anti-GFP Antibody (A11122) (Right Panel) at a 1:5,000 dilution (1 hour incubation). The lanes contain: (A) HEK293 cells transfected with GFP expressing plasmid, (B) non-transfected HEK293 cells. Detected by chemiluminescence. Discover more, for less.
