RGM-C/Hemojuvelin Antibody [Unconjugated]

RGM-C/Hemojuvelin was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human/Mouse RGM-C/Hemojuvelin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3720) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

RGM-C/Hemojuvelin Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human RGM-C isoform a (R&D Systems, Catalog # 3720-RG)
  Gln36-Asp400
  Accession # Q6ZVN8
• Specificity: Detects human and mouse RGM-C in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross‑reactivity with recombinant human (rh) RGM-B is observed and less than 5% cross-reactivity with rhRGM-A is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: HFE2
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for RGM-C/Hemojuvelin Antibody [Unconjugated]

• DL-M
• haemojuvelin
• hemochromatosis type 2 (juvenile)
• Hemojuvelin
• HFE2
• HFE2AMGC23953
• HJV
• HJVHemochromatosis type 2 protein
• JH
• repulsive guidance molecule c
• RGMC
• RGM-C
• RGMCRGM domain family member C

Background

RGM-C, also known as hemojuvelin, is a member of the repulsive guidance molecule (RGM) family of GPI-linked neuronal and muscle membrane glycoproteins (1, 2). RGM-C is expressed in striated muscle and periportal hepatocytes (3-5). The protein undergoes partial cleavage intracellularly, resulting in a disulfide-linked dimer of the 14 kDa N-terminal and 33 kDa C-terminal portions (4, 6, 7). The N-terminal fragment contains an RGD motif, while the C-terminal fragment carries the GPI attachment site (4, 7). Two alternatively spliced isoforms lack either approximately half or the entire N-terminal fragment. Full length RGM-C can also be released from the cell and circulates in the blood (6, 8). RGM-C is disrupted in type 2A juvenile hemochromatosis, a hereditary iron homeostasis disorder characterized by excessive iron accumulation (5). In mouse, loss of RGM-C function results in decreased expression of the iron regulatory hormone hepicidin and increased iron deposition in liver, pancreas, and heart (5, 9). Membrane associated RGM-C upregulates hepicidin while soluble RGM-C downregulates hepicidin expression (8). This appears to be an iron-responsive regulatory system, as high blood iron levels reduce the amount of soluble RGM-C produced (8). RGM-C, similar to RGM-A, associates with neogenin (7). Disease-related point mutations can prevent internal RGM-C cleavage or its ability to interact with neogenin (6, 7). Experimental inflammatory conditions result in decreased RGM-C expression and increased hepicidin expression, although the two effects occur independently (5, 10). RGM-C also functions as a BMP coreceptor and enhances BMP-2 and BMP-4 signaling (11). In this context, RGM-C enhances the BMP-2 upregulation of hepatic hepicidin (11). Mature human RGM-C shares 89% amino acid (aa) sequence identity with mouse and rat RGM-C. It shares 49% and 44% aa sequence identity with human RGM-A and RGM-B, respectively.

1. Papanikolaou, G. et al. (2004) Nat. Genet. 36:77.
2. Schmidtmer, J. and D. Engelkamp (2004) Gene Exp. Patterns 4:105.
3. Oldekamp, J. et al. (2004) Gene Exp. Patterns 4:283.
4. Niederkofler, V. et al. (2004) J. Neurosci. 24:808.
5. Niederkofler, V. et al. (2005) J. Clin. Invest. 115:2180.
6. Kuninger, D. et al. (2006) J. Cell Sci. 119:3273.
7. Zhang, A.S. et al. (2005) J. Biol. Chem. 280:33885.
8. Lin, L. et al. (2005) Blood 106:2884.
9. Huang, F.W. et al. (2005) J. Clin. Invest. 115:2187.
10. Krijt, J. et al. (2004) Blood 104:4308.
11. Babitt, J.L. et al. (2006) Nat. Genet. 38:531.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for RGM-C/Hemojuvelin Antibody (AF3720)(5)

Publications using AF3720

• SP Dion, F Béliveau, LP Morency, A Désilets, R Najmanovic, R Leduc Functional diversity of TMPRSS6 isoforms and variants expressed in hepatocellular carcinoma cell lines Sci Rep, 2018-08-22;8(1):12562. 2018-08-22 [PMID: 30135444] (Western Blot, Human)
• SP Dion, F Béliveau, A Désilets, MG Ghinet, R Leduc Transcriptome analysis reveals TMPRSS6 isoforms with distinct functionalities J. Cell. Mol. Med., 2018-02-14;0(0):. 2018-02-14 [PMID: 29441715] (Western Blot, Human)
• Orlando M. Gutiérrez, Chia Chi Sun, Wenjie Chen, Jodie L. Babitt, Herbert Y. Lin Statement of Concern about a Commercial Assay Used to Measure Soluble Hemojuvelin in Humans American Journal of Nephrology 2012-09-26 [PMID: 23006450]
• Wenjie Chen, Chia Chi Sun, Shanzhuo Chen, Delphine Meynard, Jodie L Babitt, Herbert Y Lin A novel validated enzyme-linked immunosorbent assay to quantify soluble hemojuvelin in mouse serum Haematologica 2013-02-01 [PMID: 22875629]
• Maegdefrau U, Arndt S, Kivorski G, Hellerbrand C, Bosserhoff AK Downregulation of hemojuvelin prevents inhibitory effects of bone morphogenetic proteins on iron metabolism in hepatocellular carcinoma. Lab. Invest., 2011-08-22;91(11):1615-23. 2011-08-22 [PMID: 21863061] (IHC-P, Western Blot, Human)

Bioinformatics

• Gene Symbol: HFE2
• Uniprot:
  • Q6ZVN8()