Anti-Green Fluorescent Protein Antibody

Article Name:	Anti-Green Fluorescent Protein Antibody
Biozol Catalog Number:	AVL-GFP-1020
Supplier Catalog Number:	GFP-1020
Alternative Catalog Number:	AVL-GFP-1020
Manufacturer:	Aves Labs
Category:	Antikörper
Application:	ELISA, ICC, IHC, WB
Immunogen:	Recombinant GFP expressed in Escherichia coli

Anti-Green Fluorescent Protein (GFP) Chicken Polyclonal Primary Antibody

The Anti-Green Fluorescent Protein (GFP) chicken polyclonal primary antibody from Aves Labs is produced in-house and detects Green Fluorescent Protein (GFP) and related variants including eGFP, YFP, CFP and others. It is a mixture of IgY fraction and affinity-purified antibodies and is great for use in ELISA, IHC, ICC, and WB.

Background

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.

Product Details

Antigen: Recombinant GFP expressed in Escherichia coli

Clonality: Polyclonal

Host Species: Chicken

Format: IgY Fraction

Concentration: 10 mg/mL

Physical State: Liquid

Molecular Weight: 27 kDa

Antibody Registry ID: AB_2307313

Applications: ELISA, ICC, IHC, WB

Dilution Ranges

WB: 1:5000-1:10000
IHC: 1:2000-1:5000
ICC: 1:2000-1:5000

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).

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.

These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.

Clonality:	Polyclonal
Concentration:	10 mg/mL
Isotype:	IgY
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.
Storage:	Store at -20°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile.
Antibody Type:	Primary Antibody

	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.
	Comparison between GFP immunoreactivity in Rexed Lamina 2 neurons of a transgenic mouse using Aves Labs’ 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.
	Immunoreactivity using anti-GFP antibody (left panel in red), Western blot showing specific immunolabeling of the GFP protein.
	Immunolabeling of dividing Drosophilia larval neuroblast using Anti-GFP (green, Cat. No. GFP-1010, 1:500) to detect the GFP tagged telomere protein HOAP, and nuclei stained with DAPI (grey). Image was taken with a Zeiss 880 confocal and kindly provided by Brandt Warecki, Sullivan Lab, University of California, Santa Cruz.
	Immunostaining of mouse hippocampus identifying injected HSV containing GFP (Cat GFP, 1:250, green) and DAPI (blue). Image kindly provided by Chad Brunswick, Penn State University.
	Immunostaining of adult hippocampal neural stem cells from Nestin GFP trangenic mice labeling GFAP (orange, mouse) and GFP (orange, 1:1000, chicken, cat co GFP). The confocal image was obtained from a Leica Sp8 confocal microscope, magnification 40X. Image kindly provided by Soraya Martín Suarez, Achucarro Basque Center for Neuroscience, Spain.
	Immunostaining of CX3CR1-Cre mouse hypothalamus showing specific detection of GFP (cat. GFP, green) and IBA1 (red). Nuclei are labeled with DAPI (blue). Image kindly provided by Karmen Ma, NIDDK – National Institutes of Health.
	Immunostaining of CX3CR1-Cre mouse hypothalamus showing specific detection of GFP (cat. GFP, green) and IBA1 (red). Image kindly provided by Karmen Ma, NIDDK – National Institutes of Health.