Mouse RGM-A Antibody

R&D Systems | Catalog # AF2458

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse RGM-A
Cys48-Gly421
Accession # Q6PCX7

Specificity

Detects mouse RGM-A in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse RGM-A Antibody

RGM-A antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

RGM‑A in Mouse Embryo.

RGM-A was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c) using Goat Anti-Mouse RGM-A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2458) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to retina and developing neurons in cross-section of the developing eye. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of RGM-A by Immunohistochemistry

Detection of RGM-A by Immunohistochemistry

Immunohistochemical analysis of the spinal cord in targeted EAE mice treated by control/anti-RGMa antibody at 14 DPI. (A) Graphical representation of IHC experiments. (B) Representative images of vascular abnormalities at 14 DPI obtained from the isotype control group. (C) Representative images of the spinal cord stained for P-selectin, type I collagen, CD31, and type IV collagen in the naïve (left), isotype control (middle), and anti-RGMa antibody groups (right) at 14 DPI. Arrows indicate abnormal vessels. Scale bar; 50 μm (b, magnified image), 250 μm (c, whole spinal cord). (D) Quantification of P-selectin, CD31, perivascular type I and IV collagen-positive areas within white matter in each group (naïve; n=8, isotype control; n=4, anti-RGMa; n=4. Error bars represent the mean ± SEM. *p < 0.05 vs naïve, #p <0.05 vs isotype control, assessed via one-way ANOVA, followed by the Tukey-Kramer test). Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2022.870126/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of RGM-A by Immunohistochemistry

Detection of RGM-A by Immunohistochemistry

Immunohistochemical analysis of the spinal cord in targeted EAE mice. (A) Experimental time course of IHC analysis. (B) Representative image of the whole spinal cord stained with anti-CD31 and anti-type IV collagen antibodies at each disease phase in naïve mice and targeted EAE mice at 8, 14, 21 DPI. (C) Representative image of anti-RGMa and anti-CD31 antibodies staining in targeted EAE mouse spinal cords at 14 and 21 DPI. White and yellow arrows indicate abnormal vessels and collagen mesh formation, respectively. Scale bar; 50 μm (magnified), 250 μm (whole spinal cord). Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2022.870126/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of RGM-A by Flow Cytometry

Detection of RGM-A by Flow Cytometry

Transcriptional analysis of targeted EAE mice treated with control/anti-RGMa antibody. (A) Graphical representation of transcriptional analysis experiments. (B) Principal component analysis plot of each group. (C) Volcano plot (left) and MA plot (right) of the comparison between the isotype control group and the anti-RGMa antibody group. Differentially expressed genes (fold change>1.5, p-value < 0.05) were represented by color dots. (D) GSEA plot of CNS cell markers with positive enrichment in anti-RGMa antibody group compared to isotype control group (p-adjust < 0.05). (E) GSEA plot of CNS cell markers with negative enrichment in anti-RGMa antibody group compared to isotype control group (p-adjust < 0.05). (F–H) Top 5 GSEA plots with negative enrichment gene sets (p-adjust < 0.05) of GO biological process (F), KEGG pathway (G), and MsigDB hall mark (H). (I) GSEA plot of BBB related genes sets (p-adjust < 0.05, n.s.; not significant). (J) Key features related to BBB disruption and corresponding genes (left) and heatmap of representative genes in BBB dysfunction modules, ECM production, and EndMT (right). Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fimmu.2022.870126/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse RGM-A Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed frozen sections of mouse embryo (13 d.p.c.)

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse Repulsive Guidance Molecule A/RGM-A (Catalog # 2458-RG)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: RGM-A

Mouse repulsive guidance molecule (RGM-A) is a 33 kDa GPI-linked member of an expanding RGM-related family of neuronal and muscle-expressed membrane proteins (1). It is synthesized as a 454 amino acid (aa) preproprotein that contains a 47 aa signal sequence, a 122 aa N-terminal prosegment, a 258 mature region and a 27 aa C-terminal prosegment (2, 3). The N-terminal prosegment contains an RGD tripeptide and the molecule’s only two potential N-linked glycosylation sites. The mature segment shows an abbreviated von Willebrand factor domain. Proteolytic processing occurs at an aspartic acid-proline bond, creating a predicted 32 kDa mature region (2). The mature region of mouse RGM-A is 93% and 87% aa identical to human and chick mature region RGM-A, respectively. When compared to mouse RGMb and c, the mature region of mouse RGM-A shows 77% and 76% aa identity, respectively (2, 3). Recombinant chick RGM has been reported to induce collapse of temporal but not nasal growth cones, and to repel temporal retinal axons in vitro. This suggests a role in the development of the retina-superior colliculus connection. In mice, however, this activity is not so obvious, and thus its function in this system is uncertain (3). Alternatively, in mice, RGM-A is said to be needed for neural tube closure (3). And in a mouse culture system, chick RGM-A is reported to be responsible for the layered segmentation of of entorhinal cortical projections to the hippocampus (4). The receptor for RGM is reported to be neogenin (5, 6).

References

  1. Samad, T.A. et al. (2004) J. Neurosci. 24:2027.
  2. Schmidtmer, J. et al. (2004) Gene Expr. Patterns 4:105.
  3. Niederkofler V. et al. (2004) J. Neurosci. 24:808.
  4. Brinks, H. et al. (2004) J. Neurosci. 24:3862.
  5. Rajagopalan S. et al. (2004) Nat. Cell Biol. 6:756.
  6. Matsunaga E. et al. (2004) Nat. Cell Biol. 6:749.

Long Name

Repulsive Guidance Molecule A

Alternate Names

RGMA

Entrez Gene IDs

56963 (Human); 244058 (Mouse)

Gene Symbol

RGMA

UniProt

Additional RGM-A Products

Product Documents for Mouse RGM-A Antibody

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Product Specific Notices for Mouse RGM-A Antibody

For research use only

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Citations for Mouse RGM-A Antibody

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Protocols

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