Nephronectin Antibody [Unconjugated]

Nephronectin was detected in immersion fixed paraffin-embedded sections of mouse pancreas using Goat Anti-Mouse Nephronectin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4298) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to islet cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Mu
• Applications: IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Nephronectin Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Nephronectin
  Asp20-Cys561
  Accession # AAH68308
• Specificity: Detects mouse Nephronectin in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: NPNT
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 1-15 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 Nephronectin Antibody [Unconjugated]

• EGFL6L
• Nctn
• Nephronectin
• NPNT
• POEM

Background

Nephronectin, also named POEM (preosteoblast EGF repeat protein with MAM domain) is a 70‑90 kDa extracellular matrix protein that is a ligand for integrin alpha ₈ beta ₁ (1, 2). The 561 amino acid (aa) mouse Nephronectin (isoform b) contains a 19 aa signal sequence, a matrilin-type vWA domain, two calcium-binding EGF-like domains, a potential N-linked glycosylation site, a pro/ser/thr-rich mucin-like region, an RGD integrin binding motif, and a MAM domain. These features are often present in matrix proteins that, like Nephronectin, mediate cell adhesion and spreading (1, 2). Isoform a includes a 17 aa insertion between aa 58 and 59 that is missing in isoform b. Mouse Nephronectin shows 88%, 92%, 85% and 77% aa identity with human, rat, bovine and opossum Nephronectin, respectively. It is most highly expressed in developing endocrine organs such as parathyroid, thyroid, hypophysis and pineal organ, and around developing bone, teeth, and muscle (1, 2). Pre-osteoblast cells produce Nephronectin, but down‑regulate it as they differentiate (1). It is also expressed in the Wolffian duct and ureteric bud basement membranes in the developing kidney, where it is colocalizes, and forms an in-vivo complex with integrin alpha ₈ beta ₁ (2). Since deletion of integrin alpha ₈ beta ₁ results in renal agenesis, Nephronectin has been proposed to be a critical integrin alpha ₈ beta ₁ ligand during Nephrogenesis (2, 3).

1. Morimura, N. et al. (2001) J. Biol. Chem. 276:42172.
2. Brandenberger, R. et al. (2001) J. Cell Biol. 154:447.
3. Miner, J. H. (2001) J. Cell Biol. 154:257.

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 Nephronectin Antibody (AF4298)(6)

Publications using AF4298

• J Ma, L Bi, J Spurlin, P Lwigale Nephronectin-integrin alpha8 signaling is required for proper migration of periocular neural crest cells during chick corneal development Elife, 2022-03-03;11(0):. 2022-03-03 [PMID: 35238772] (Stimulation, Chicken)
• J Su, U Sabbagh, Y Liang, L Olejníková, KG Dixon, AL Russell, J Chen, YA Pan, JW Triplett, MA Fox A cell-ECM mechanism for connecting the ipsilateral eye to the brain Proceedings of the National Academy of Sciences of the United States of America, 2021-10-19;118(42):. 2021-10-19 [PMID: 34654745] (IHC, Mouse)
• Yulia Shwartz, Meryem Gonzalez-Celeiro, Chih-Lung Chen, H. Amalia Pasolli, Shu-Hsien Sheu, Sabrina Mai-Yi Fan et al. Cell Types Promoting Goosebumps Form a Niche to Regulate Hair Follicle Stem Cells Cell 8/1/2020 [PMID: 32679029]
• Alicia Bellomo, Isabelle Mondor, Lionel Spinelli, Marine Lagueyrie, Benjamin J. Stewart, Nicolas Brouilly et al. Reticular Fibroblasts Expressing the Transcription Factor WT1 Define a Stromal Niche that Maintains and Replenishes Splenic Red Pulp Macrophages Immunity 7/1/2020 [PMID: 32562599]
• Kato T, Yamada A, Ikehata M et al. FGF-2 suppresses expression of nephronectin via JNK and PI3K pathways FEBS Open Bio 5/11/2018 [PMID: 29744297]
• Tamaki Kurosawa, Atsushi Yamada, Dai Suzuki, Naoko Morimura, Yoshiyuki Sasagane, Hiroyuki Itabe et al. Nephronectin Expression Is Up-Regulated by BMP-2 Biological & Pharmaceutical Bulletin 7/5/2016 [PMID: 27374296]

Bioinformatics

• Gene Symbol: NPNT
• Uniprot:
  • AAH68308()