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Key Documents

ABN1723

Sigma-Aldrich

Anti-RTN3 (R458)

from rabbit

Synonyme(s) :

Reticulon-3, HAP, Homolog of ASY protein, Neuroendocrine-specific protein-like 2, Neuroendocrine-specific protein-like II, NSP-like protein 2, NSP-like protein II, NSPLII

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About This Item

Code UNSPSC :
12352203
eCl@ss :
32160702
Nomenclature NACRES :
NA.41

Source biologique

rabbit

Niveau de qualité

Forme d'anticorps

unpurified

Type de produit anticorps

primary antibodies

Clone

polyclonal

Espèces réactives

mouse, human

Réactivité de l'espèce (prédite par homologie)

rat (based on 100% sequence homology), chimpanzee (based on 100% sequence homology)

Technique(s)

immunofluorescence: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

Numéro d'accès NCBI

Numéro d'accès UniProt

Conditions d'expédition

dry ice

Modification post-traductionnelle de la cible

unmodified

Informations sur le gène

human ... RTN3(10313)

Description générale

Reticulon-3 (UniProt O95197; also known as HAP, Homolog of ASY protein, Neuroendocrine-specific protein-like 2, Neuroendocrine-specific protein-like II, NSP-like protein 2, NSP-like protein II, NSPLII) is encoded by the RTN3 (also known as ASYIP, NSPL2) gene (Gene ID 10313) in human. Reticulon-3/RTN3 belongs to the RTN family of proteins characterized by their C-terminal reticulon homolog domain (RHD; a.a. 844-1032 in RTN3). There exist four mammalian reticulons (RTN1-4), with each member having multiple spliced isoforms. RTN3 is expressed in neurons and localized in axons and growth cones. RTN3 overexpression in neurons results in reduced amyloid deposition in murine models of Alzheimer′s disease (AD). Consistently, RTN3 deficiency elevates BACE1 protein level and enhances amyloid deposition in AD mice that express human mutant APP and presenilin-1 transgenes. Human RTN3 is a multipass ER and golgi membrane protein with 3 membrane helical regions (a.a. 864-997, 948-968, 973-993) and four cytoplasmic regions (a.a. 2-863, 888-947, 969-972, 994-1032). Its C-terminal RHD mediates interaction with FADD (a.a. 987-1032) and BACE1 (a.a.1000-1002). Alternative splicings generate 7 isoforms (UniProt O95197-1 througth O95197-7).

Spécificité

Target specificity of this rabbit polyclonal antiserum has been verified by Western blotting analysis of RNAi-mediated RTN3-knockdown in Swedish mutant APP-expressing 125.3 cells and tissue samples from RTN3-knockout mice. Immunogen sequence is located near the C-terminus of all human/mouse/rat isoforms reported by UniProt (O95197, O95197, Q9ES97) with the exception of human isoforms 5 and 6 (O95197-5 and O95197-6), which lack the target sequence.

Immunogène

Conjugated linear peptide corresponding to a sequence near the C-terminus of human RTN3.

Application

Anti-RTN3 (R458) Antibody, Cat. No. ABN1723, is a highly specific rabbit polyclonal antibody that targets RTN3 and has been tested in and Immunofluorescence, Immunohistochemistry, Immunoprecipitation, and Western Blotting.
Immunofluorescence Analysis: A 1:2,000 dilution from a representative lot immunostained dystrophic neurites in Alzheimer′s diseased (AD) human frozen brain tissue sections by fluorescent immunohistochemistry (Courtesy of Wanxia He, The Cleveland Clinic Foundation, U.S.A.).

Immunohistochemistry Analysis: A 1:2,000 dilution from a representative lot immunostained dystrophic neurites in Alzheimer′s diseased (AD) human frozen brain tissue sections (Courtesy of Wanxia He, The Cleveland Clinic Foundation, U.S.A.).

Western Blotting Analysis: A 1:2,000 dilution from a representative lot detected RTN3 isoforms in 50 µg of brain tissue lysate from a wild-type, but not RTN3-knockout, mouse (Courtesy of Wanxia He, The Cleveland Clinic Foundation, U.S.A.).

Immunofluorescence Analysis: A representative lot detected RTN3 immunoreactivity partially co-localized with that of BACE1 in mouse brain tissue sections (He, W., et al. (2004). Nat. Med.10(9):959-965).

Immunohistochemistry Analysis: A representative lot detected RTN3 immunoreactivity enriched in grey matter neuronal cell bodies of human brain tissue sections (He, W., et al. (2004). Nat. Med.10(9):959-965).

Immunoprecipitation Analysis: A representative lot co-immunoprecipitated BACE1 with RTN3 from HEK293 and human frontal cortex membrane extracts (He, W., et al. (2004). Nat. Med.10(9):959-965).

Western Blotting Analysis: A representative lot detected RTN3 isoforms expression in multiple mouse tissues, including broadly expressed ~25 kDa RTN3-A1 (UniProt Q9ES97-3), ~100 kDa fat tissue-specific RTN3-AL (UniProt Q9ES97-1), ~95 kDa brain-specific RTN3-B, and ~27 kDa RTN3-A2 (UniProt Q9ES97-4) in spinal cord. RTN3 target bands were not detected in tissue samples from RTN3-knockout mice (Shi, Q., et al. (2014). J. Neurosci. 34(42):13954-13962; He, W., et al. (2004). Nat. Med.10(9):959-965).

Western Blotting Analysis: A representative lot detected RTN3 isoform 3 (UniProt O95197-3) wild-type and mutant constructs expression. Membrane integration was affected by L71K/L72K mutation or deletion of N-terminal 97 a.a., while truncation of the first 61 a.a. did not affect embrane integration. Western blotting analysis of protease K-digested HEK293 microsomal preparation indicated a cytoplasmic orientation of the C-terminal end (He, W., et al. (2007). J. Biol. Chem. 282(40):29144-29151).

Western Blotting Analysis: A representative lot detected RTN3 in BACE1 immunoprecipitate from HEK293 membrane extract. A bimodal distribution showing RTN3 enrichment in subcellular fractions containing Golgi proteins and those containing ER markers was seen (He, W., et al. (2004). Nat. Med.10(9):959-965).

Western Blotting Analysis: A representative lot detected RNAi-mediated RTN3-knockdown in Swedish mutant APP-expressing 125.3 cells (He, W., et al. (2004). Nat. Med.10(9):959-965).
Research Category
Neuroscience

Qualité

Evaluated by Western Blotting in mouse brain tissue lysates.

Western Blotting Analysis: A 1:500 dilution of this antibody detected RTN3 isoforms in 50 µg of brain tissue lysate from a wild-type, but not RTN3-knockout, mouse.

Description de la cible

~95/25 kDa observed. 112.5/110.5/25.48/27.44/100.7 kDa (human isoform 1/2/3/4/7), 103.7/101.8/25.30/27.26/68.64 kDa (mouse isoform 1/2/3/4/5), 101.4/25.30 kDa (rat isoform 1/2) calculated. Uncharacterized bands may be observed in some lysate(s).

Forme physique

Format: Unpurified
Rabbit polyclonal antibody serum with 0.05% sodium azide.
Unpurified.

Stockage et stabilité

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Autres remarques

Concentration: Please refer to lot specific datasheet.

Clause de non-responsabilité

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 1

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


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Consulter la Bibliothèque de documents

Youtong Huang et al.
Nature immunology, 22(5), 586-594 (2021-04-17)
Two microglial TAM receptor tyrosine kinases, Axl and Mer, have been linked to Alzheimer's disease, but their roles in disease have not been tested experimentally. We find that in Alzheimer's disease and its mouse models, induced expression of Axl and
Tarique R Bagalkot et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 41(2), 234-250 (2020-11-26)
Dopamine transporter (DAT) controls dopamine neurotransmission by clearing synaptically released dopamine. However, trafficking itineraries of DAT, which determine its cell-surface concentration near synapses, are poorly characterized. It is especially unknown how DAT is transported between spatially distant midbrain somatodendritic and

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