Anti-MAGE-D1 / Dlxin-1 / NRAGE antibody, rabbit serum (MD1)
Product code74-112Size

100 µlStorage

Store at 4℃ for short term. For long term storage store at -20℃.
Aliquot to avoid repeated freezing and thawing.

Concentration

N/A

BufferPreservative: 0.05% sodium azide

PurityRabbit antiserum

Immunogen

Recombinant MBT-fused mouse MAGE-D1 (aa 1-775 ).

IsotypeRabbit IgG

Reactivity

Mouse, rat and human MAGE-D1

Special notes

N/A

Application

1. Western blotting (dilution: 1/3,000-1/1,000)
2. Immunohistochemistry (dilution: 1/300)
3. Immunocytochemistry (dilution: 1/1,000-1/500)
4. ELISA Not tested for other applications

Background

MAGE-D1 (melanoma-associated antigen D1), also known as Dlxin-1 orNRAGE, is a member of the MAGE protein family. MAGE-D1 is expressed inalmost all normal adult tissues and has been demonstrated to interact with thep75 neutrophin receptor and to facilitate nerve growth factor-dependent
apoptosis (ref.1). MAGE-D1 also interacts with its homologous protein, necdin,
which is known as a growth suppressor and a promoter of differentiation inneurons. Necdin binds to Msx (msh homeobox) and Dlx (distal-less homeobox)
family homeodomain transcription factors via MAGE-D1. These proteins
cooperate to modulate differentiation of neurons and muscle cells by regulating
gene expression whereas Msx proteins function as transcriptional repressor andDlx proteins function as transcriptional activators (ref.2 and 3).
An antibody (named MD1) against mouse MAGE-D1 was raised in rabbit (ref.2).

Data Link

UniProtKB Q9QYH6 (mouse), Q9ES73 (rat), Q9Y5V3 (human)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 74-112 anti-MAGE-D1 / Dlxin-1 / NRAGE antibody, rabbit serum (MD1)

Fig.1 Western blot analysis of MAGE-D1 expression in various organs in mouse embrio.
Tissue lysates (20 µg) from E18.5 embryos were separated by SDS-PAGE and immunoblotted withantibodies against necdin, MAGE-D1 (D1) and tubulin.
Endogenous ~85-kDa MAGE-D1 protein was expressed in a ubiquitous manner.
Tubulin was used as internal control.

Fig.2.Western blotting of MAGE-D1 in the crude extract of mouse embryo.
The extract (10µg protein) was prepared from cerebral cortex of E 16.5 mouse embryo. The antiMAGE-D1 antibody (MD1) was used at 1/3,000 dilution. Molecular mass of mouse MAGE-D1is 86kDa. The numbers are positions of marker proteins shown in kDa.

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Fig.2 Immunohistochemistry for MAGE-D1 in mouse embryos.
Cryostat section of the paraformaldehyde fixed forebrain at E12.5 embryo (upper panel) and the hindlimb at E14.5 (lower panel) were stained with antibody against MAGE-D1 at 1/300 dilution.
The arrows point to the preplate (upper panel) and the skeletal muscles (lower panel). The
arrowheads indicate the ventricular proliferative zone; V, ventricle; C, bone cavity.
MAGE-D1 was concentrated in the preplate of the forebrain at E12.5 and skeletal muscle tissues in
the hind limb at E14.5. In developing neural tube, MAGE-D1 immunoreactivity was distributed in
the ventricular zone as well as the marginal zone.

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Fig.3 Immunocytochemistry using this antibody .
C2C12 myoblast cells were cultured under undifferentiated conditions (UD) or under differentiation
conditions for 4 days (4D). Cells were fixed and triply stained for myosin heavy chain (MHC) withanti-MHC antibody (non BioAcademia product) and for MAGE-D1 (D1) with anti-MAGE-D1 antibody,and for nuclear DNA with Hoechst33342 (Hoechst). The arrowheads point to differentiatedmultinucleated myocytes.
MHC and MAGE-D1 were distributed predominantly in the cytosol of multinucleated differentiated
C2C12 cells.
Related product:74-100 anti-Necdin antibody.
74-114 anti-MAGE-G1 / Necdin-like 2 antibody
References: This antibody was produced and used in ref.2 and 3.
1. Salehi AH et al (2000) “NRAGE, a novel MAGE protein, interacts with the p75 neutrophin receptor
and facilitates nerve growth factor dependent apoptosis.” Neuron 27: 279-288 PMID: 10985348
2. Kuwajima T et al (2004) “Necdin interacts with the Msx2 homeodomain protein via MAGE-D1 topromote myogenic differentiation of C2C12 cells.” J Biol Chem 279: 40484-40493 PMID: 15272023
3. Kuwajima T et al (2006) “Necdin promotes GABAergic neuron differentiation in cooperation withDlx homeodomain proteins.” J Neurosci 26: 5383-5392 PMID: 16707790

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