Staining of a tissue section through the midbrain region of an adult mouse for GFP (green) and Tryptophan Hydroxylase-positive neurons (RED).

Staining of a tissue section through the midbrain region of an adult mouse for GFP (green) and Tryptophan Hydroxylase-positive neurons (RED).

Pyramidal neurons in the hippocampal formation of a neonatal mouse brain. Tissue was paraformaldehyde-fixed (4%) and paraffin-embedded. GFP staining is in green in left panel.

Pyramidal neurons in the hippocampal formation of a neonatal mouse brain. Tissue was paraformaldehyde-fixed (4%) and paraffin-embedded. GFP staining is in green in left panel.

Comparison between GFP immunoreactivity in Rexed Lamina 2 neurons of a transgenic mouse using AVESLABS anti-GFP (GREEN) and 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 in Rexed Lamina 2 neurons of a transgenic mouse using AVESLABS anti-GFP (GREEN) and 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.

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.

from Aves Labs

Anti-Green Fluorescent Protein Antibody (GFP)

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Product Details | ↓ Citations



Volume: 100 µL (GFP-1010) or 400 µL (GFP-1020)

Concentration: 10 mg/mL

Clonality: Polyclonal

Form: IgY Fraction

Host Species: Chicken

Applications: ELISA, ICC, IHC, WB

Target Description: Green Fluorescent Protein (GFP) is a naturally fluorescent protein originally derived from jellyfish. GFP has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors which have become useful and ubiquitous tools in transgenic experiments. Fluorescent proteins enable a wide range of applications where they have functioned as cell lineage tracers, reporters of gene expression, or as a measure of protein-protein interactions.

Antibody Registry ID (RRID): AB_2307313

Physical State: Liquid

Buffer: Sodium phosphate (10 mM, pH 7.2) buffered isotonic saline (0.9%, w/v), glycerol (50%, v/v), with sodium azide (0.02%, w/v) as an anti-microbial agent.

Production Notes: 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).

Validation and Application Notes

Western Blot Dilution Range: 1:5000-1:10000

IHC Dilution Range: 1:2000-1:5000

Quality Control

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.

Storage

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.

NOTE

Aves Labs products are intended for use as research laboratory reagents. They are not intended for use as diagnostic or therapeutic reagents in humans.


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