F-Spondin/SPON1 Antibody [Unconjugated]

Western blot shows lysates of OVCAR-3 human ovarian carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human F-Spondin/SPON1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3135) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for F-Spondin/ SPON1 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of OVCAR‑3 human ovarian carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for F‑Spondin/SPON1 at approximately 134 kDa (as indicated) using 10 µg/mL of Goat Anti-Human F‑Spondin/SPON1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3135) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

F-Spondin/SPON1 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human F‑Spondin/SPON1
  Phe29-Cys807
  Accession # Q9HCB6
• Specificity: Detects human F-Spondin/SPON1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SPON1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Neutralization 3 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 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 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC 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 F-Spondin/SPON1 Antibody [Unconjugated]

• FSpondin
• F-Spondin
• KIAA0762VSGP/F-spondin
• MGC10724
• SPON1
• spondin 1, (f-spondin) extracellular matrix protein
• spondin 1, extracellular matrix protein
• spondin-1
• Vascular smooth muscle cell growth-promoting factor
• VSGP

Background

F-Spondin (“Floor plate” and “thrombospondin” homology; also Spondin-1 and VSGP) is a member of the F-Spondin family of proteins that collectively belong to a subgroup of TSR (thrombospondin) type I class molecules (1). Class I molecules are either membrane-bound or ECM-associated. F-Spondin is a 110 kDa, secreted, heparin-binding extracellular matrix glycoprotein first characterized in rat for its high expression in embryonic floor plate (2-4). Human F-Spondin is synthesized as an 807 amino acid (aa) precursor that contains a 28 aa signal sequence and a 779 aa mature region (3, 4). The mature region includes an N-terminal reelin-like domain (aa 1-200), a centrally placed F-Spondin (FS) type segment (aa 201-440), and six C-terminal class 2 thrombospondin type I repeats (1, 3, 5). Class 1 and 2 repeats differ in the placement of their cysteine residues. The fifth and sixth TSP repeats (aa 668-806) apparently bind ECM, while TSP repeats 1-4 (aa 442-666), plus the spondin segment, are suggested to mediate either repulsive activity (on motor neurons), or outgrowth promoting activity (on sensory neurons) (1, 6). At least two isoforms of F-Spondin are known. Both are proteolytically-generated, one by plasmin, another by an unidentified protease. Plasmin cleaves the C-terminus at two points, generating a soluble, 95 kDa, 656 aa F-Spondin that contains all but TSP repeats #5 and 6 (7). The unidentified protease appears to cleave F-Spondin between the FS segment and the first TSP repeat, generating 60 kDa and 50 kDa fragments, respectively (6). F-Spondin shows highly unusual glycosylation, exhibiting both C-mannosylation (mannose bound to Trp) and O-fucosylation (fucose bound to Ser/Thr) (4). The significance of these glycosidic modifications is unknown. Mature human F-Spondin is 98%, 97%, 98%, and 97% aa identical to mature canine, rat, bovine and mouse F-Spondin, respectively. Mammalian cells known to express F-Spondin include floor plate epithelium, ventral motor neurons, Schwann cells, fibroblasts, hippocampal pyramidal cells, endothelial cells, vascular smooth muscle cells and some tumor cells (6, 8, 9).

1. Feinstein, Y. and A. Klar (2004) Int. J. Biochem. Cell Biol. 36:975.
2. Klar, A. et al. (1992) Cell 69:95.
3. Miyamoto, K. et al. (2001) Arch. Biochem. Biophys. 390:93.
4. Gonzalez de Peredo, A. et al. (2002) Mol. Cell. Proteomics 1:11.
5. Adams, J.C. and R.P. Tucker (2000) Dev. Dyn. 218:280.
6. Burstyn-Cohen, T. et al. (1998) J. Neurosci. 18:8875.
7. Tzarfaty-Majar, V. et al. (2001) J. Biol. Chem. 276:28233.
8. Feinstein, Y. et al. (1999) Development 126:3637.
9. Pyle-Chenault, R.A. et al. (2005) Tumor Biol. 26:245.
10. Mattes, J. and P.S. Foster (2003) Curr. Drug Targets Inflamm. Allergy 2:169.

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 F-Spondin/SPON1 Antibody (AF3135)(2)

Publications using AF3135

• Ramiro Rincón-Rodriguez, Dennise Mena, Javier Mena, Patricia Díaz-Saldivar, Emanuel Guajardo-Correa, Carlos Godoy-Guzman et al. F-Spondin Is the Signal by Which 2-Methoxyestradiol Induces Apoptosis in the Endometrial Cancer Cell Line Ishikawa International Journal of Molecular Sciences 8/7/2019 [PMID: 31394756]
• J Song, SL Merbs, LJ Sokoll, DW Chan, Z Zhang A multiplex immunoassay of serum biomarkers for the detection of uveal melanoma Clin Proteomics, 2019-03-05;16(0):10. 2019-03-05 [PMID: 30867659] (Human)

Bioinformatics

• Gene Symbol: SPON1
• Uniprot:
  • Q9HCB6()