Photomicrograph of a paraffin-embedded tissue section through an adult dentate gyrus of the hippocampal formation from a paraformaldehyde-fixed (4%) mouse brain. RED shows nestin immunoreactivity (1:1000) as visualized with a Texas Red goat anti-chicken IgY antibody (Aves Labs, 1:500). Green is staining of the granule cells; BLUE is DAPI nuclear staining. Page Balich, Univ. Arizona.
Photomicrograph of 3T3 cells in culture. Nestin immunoreactivity (RED staining, Aves Labs, 1:1000 dilution; GREEN is -Tubulin 3 staining using a rabbit antibody (Rockland, 1:500 dilution); BLUE is DAPI nuclear staining. Page Balich, Univ. Arizona.
Immunocytochemistry showing nestin immunoreactivity (GREEN; 1:1000 dilution) in A7r5 neuroblastoma cells in culture. BLUE is DAPI nuclear staining. Page Balich, Univ. Arizona.
Photomicrograph of A7r5 neuroblastoma cells in culture. Nestin immunoreactivity (RED staining, Aves Labs, 1:1000 dilution; GREEN is -Tubulin 3 staining using a rabbit antibody (Rockland, 1:500 dilution); BLUE is DAPI nuclear staining. Page Balich, Univ. Arizona.
Photomicrograph of a tissue section through the periventricular zone of an e16 mouse brain (4.0% paraformaldehyde-fixed). GREEN is nestin immunoreactivity (Aves Labs, 1:1000 dilution). BLUE is DAPI nuclear staining. Hoda Ilias, Aves Labs.
"Nestin Western Blot. 20 ug of total protein from adult mouse brain in each lane. Lane A is Aves Labs' antibody (1:2000), showing a single, strong band at the correct molecular weight. Lane B uses a rabbit antibody from BD Transduction Labs showing a weak band at the same molecular weight, as well as another spurious band at about 145 kDa. Hoda Ilias, Aves Labs."
Three antipeptide antibodies were generated in chickens against a sequence in mouse (NP_057910) gene product. Antibodies were affinity-purified and mixed in equal concentration. These antibodies have been validated with human, mouse and rat tissues.
Volume: 1000 µL
Concentration: 300 µg/mL
Host Species: Chicken
Species Reactivity: Mouse, Rat
Applications: ICC, IHC, WB
Protein Name / Synonyms: Nestin
Target Description: Mouse Nestin is a 206,994 dalton protein (1864 amino acids) expressed in neural crest cells, CNS neural stems cells, as well as a few non-neural cell types in the embryo, including cells within the pancreatic islets of Langerhans and the limb bud. Nestin is an intermediate filament-associated protein, and contributes to the cytoskeleton.
Gene ID: NES Nbla00170
Antibody Registry ID (RRID): AB_2314882
Physical State: Liquid
Production Notes: Chickens were immunized with three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the mouse nestin gene product (NM_NP_057910), but are shared with the rat (NM_012987, NCBI) protein sequence. After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, and the concentrations of the eluates adjusted to 300 µg/mL. Finally, equal volumes of each of the three affinity-purified anti-peptide antibodies were mixed, and the preparation was filtersterilized.
Validation and Application Notes
Molecular Weight: 250 kDa
Western Blot Dilution Range: 1:10000-1:20000
IHC Dilution Range: 1:1000-1:2000
Each of the three antibodies were analyzed by immunohistochemistry (at a dilution of 1:20000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.
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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