Source: https://www.aveslab.com/collections/epitope-tag-6xhis-beta-gal-actin-and-gfp-antibodies/products/green-fluorescent-protein-gfp-antibody
Timestamp: 2019-04-24 09:49:15+00:00

Document:
Anti-Green Fluorescent Protein Antibody Anti-Green Fluorescent Protein Antibody – Aves Labs, Inc.
Co-localization of GFP immunoreactive neurons (GREEN) and Tryptophan Hydroxylase-positive neurons (RED) in a tissue section through the midbrain region of an adult mouse. It isn't clear what caused the GFP-positive cells to express GFP immunoreactivity.
Pyramidal neurons in the hippocampal formation of a neonatal mouse brain. Tissue was paraformaldehyde-fixed (4%) and paraffin-embedded. GFP is the GREEN staining at the left. It isn't clear what the RED staining at the right is. The center panel shows GFP (GREEN) and this other staining (RED), plus DAPI staining (BLUE).
Comparison between GFP immunoreactivity in Rexed Lamina 2 neurons of a transgenic mouse using Aves' chicken anti-GFP (GREEN) and AbCam's rabbit anti-GFP (RED). The transgenic mouse was generated by placing the GFP cDNA after a POMC gene promoter in the transfected plasmid. Mark Zilka, Univ. North Carolina.
Comparison between GFP-immunoreactivity using Aves' anti-GFP antibody (LEFT panel in RED) and autofluorescence (RIGHT panel in GREEN). In this case, the cortical neuron in this unfixed thick section was first photographed for GFP autofluorescence (LEFT), and then the section was fixed (4% paraformaldehyde) and immunostained for GFP-immunoreactivity (1:1000 dilution) using Texas Red-goat anti-chicken IgY antibodies (Jackson ImmunoResearch) as a secondary. The same cell (LEFT) was then identified.
Chickens were immunized with purified recombinant green fluorescent protein (GFP) emulsified in Freund’s adjuvant. After multiple injections, eggs were collected from the hens, and IgY fractions were prepared from the yolks and then affinity-purified antibodies were prepared using GFP conjugated to an agarose matrix. The final product is a filter-sterilized mixture of both affinity-purified antibodies (30 µg/mL) and purified IgY (10 mg/mL).
Antibodies were analyzed by western blot analysis (1:5000 dilution) and immunohistochemistry (1:500 dilution) using transgenic mice expressing the GFP gene product. Western blots were performed using BlokHen® (Aves Labs) as the blocking reagent, and HRP-labeled goat anti-chicken antibodies (Aves Labs, Cat. #H-1004) as the detection reagent. Immunohistochemistry used tetramethyl rhodamine-labeled anti-chicken IgY.
Store at -20°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Since 50% glycerol is present in the vial, this antibody preparation should remain a liquid at -20°C. For longer storage periods, store at -80°C, but be aware that freezing this preparation may reduce its activity.
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