The immunogen is a Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246aa) derived from the jellyfish Aequorea victoria.
Conjugation:
Unconjugated
Alternative Names:
chicken anti-GFP antibody, chicken Anti-Green Fluorescent Protein Antibody, GFP antibody, Green Fluorescent Protein antibody, EGFP, enhanced Green Fluorescent Protein, Aequorea victoria, Jellyfish antibody
Clonality:
Polyclonal
Concentration:
1.18 mg/ml by UV absorbance at 280 nm
Buffer:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Form:
Liquid (sterile filtered)
Antibody Type:
Primary Antibody
Application Dilute:
ELISA: 1:2,000 - 1:12,000, IHC: 1:200 - 1:1,000, IF Microscopy: User Optimized, WB: 1:200 - 1:1,000
Application Notes:
Polyclonal anti-GFP is designed to detect GFP and its variants. Anti-GFP Antibody has been tested by ELISA, dot blot, and western and is suitable in immunofluorescence. This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct
Western Blot of anti-GFP antibody. Lane 1: mouse spleen lysate (p/n W10-000-T029). Lane 2: mouse spleen lysate spiked with 50 ng of wt GFP (p/n W10-000-T029/000-001-215). Load: 20 µg per lane. Primary antibody: GFP antibody at 2 µg/ml for 2 hr at room temperature. Secondary antibody: IRDye(TM)800 Conjugated Affinity Purified anti-Chicken IgG [H&L] [Goat] MX10 (p/n 603-132-126) at 1:20,000 for 45 min at RT. Block: 5% BSA in PBS 2 hr at room temperature. Predicted/Observed size: 27 kDa for GFP epitope tag. Other band(s): none.
Immunofluorescence Microscopy of chicken anti-GFP antibody. Tissue: KruppleGAL4 driver line in Drosophila eye disc. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: anti-GFP antibody diluted 1:500 for 2 hr at RT. Secondary antibody: Alexa(TM)488 conjugated anti-Chicken IgG at 1:300 for 1 hr at RT. Blocking: 5% NGS in PBS with 0.1% Triton X-100 for 15 min. Staining: recombinant tau-myc-GFP protein as green fluorescent signal.
Western Blot Results of Chicken Anti-GFP Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: GFP (p/n 000-001-215) / HeLa WCL (p/n W09-000-364) [0.05µg/10µg]. Lane 3: GFP (p/n 000-001-215) / NIH3T3 WCL (p/n W10-000-371) [0.05µg/10µg]. Lane 4: GFP (p/n 000-001-215) / PC-12 WCL (p/n W120-001-GL9) [0.05µg/10µg]. Lane 5: HeLa WCL (p/n W09-000-364) [10µg]. Lane 6: NIH3T3 WCL (p/n W10-000-371) [10µg]. Lane 7: PC-12 WCL (p/n W120-001-GL9) [10µg]. Primary Antibody: Anti-GFP at 1.0µg/mL overnight at 2-8C. Secondary Antibody: Goat Anti-Chicken IgG HRP (p/n 603-103-126) at 1:40,000 for 30 mins at RT. Block: MB-070 Buffer for 1hr at RT. Predicted MW: 27kda. Exposure: 2 seconds.
Western Blot of Chicken anti-GFP Antibody with Serums. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: GFP/Human Serum (p/n 000-001-215/D314-05) [0.01/0.02µL] [+]. Lane 3: Human Serum (p/n D314-05) [0.02µL] [-]. Lane 4: GFP/Mouse Serum (p/n 000-001-215/D308-05) [0.01/0.02µL] [+]. Lane 5: Mouse Serum (p/n D308-05) [0.02µL] [-]. Lane 6: GFP/Rat Serum (p/n 000-001-215/D310-05) [0.01/0.02µL] [+]. Lane 7: Rat Serum (p/n D310-05) [0.02µL] [-]. Primary antibody: Anti-GFP antibody at 1.0ug/mL overnight at 2-8C. Secondary antibody: Donkey Anti-Chicken IgG HRP secondary antibody (p/n 603-103-126) at 1:40,000 for 30 min at RT. Blocking Buffer: BlockOut Buffer (p/n MB-073) for 1hr at RT. Predicted/Observed size: ~27kDa for GFP.
Western Blot of Anti-GFP (CHICKEN) antibody. Lane 1: MW. Lane 2: GFP (p/n 000-001-215). Load: 0.05 µg. Primary antibody: Anti-GFP (CHICKEN) antibody (p/n 600-901-215) was used at 1:1000 overnight at 4C. Secondary antibody: Anti-Chicken IgG (GOAT) peroxidase conjugated antibody (p/n 603-103-126) secondary antibody was used at 1:40,000 in Blocking Buffer for Fluorescent Western Blotting (p/n MB-070). Block: 1% BSA-TTBS (p/n MB-013, diluted to 1X) 30 min at 20C. Predicted/Observed size: 27 kDa for GFP. Other band(s): none.
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