Recombinant A. caninum NIF Protein, CF

• Reactivity: Ac
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant A. caninum NIF Protein, CF Summary

• Details of Functionality: Measured by its ability to inhibit the adhesion of PMA-activated human neutrophils. The ED₅₀ for this effect is 0.1-0.4 μg/mL. Also measured by its ability to bind Recombinant Human Integrin alpha M beta 2 (Catalog # 4047-AM) in a functional ELISA.
• Source: Mouse myeloma cell line, NS0-derived a. caninum NIF protein
  Asn18-Leu274, with a C-terminal 6-His tag
• Accession #: AAA27789
• N-terminal Sequence: Asn18
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 30 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 55-65 kDa, reducing conditions

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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, and DTT.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 200 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant A. caninum NIF Protein, CF

• NIF

Background

Neutrophil inhibitory factor (NIF) of the dog hookworm, Ancylostoma caninum, is a 41 kDa secreted glycoprotein that is thought to allow the hookworm to evade the innate immune defenses of the host (1, 2). NIF interacts with the leukocyte integrin alpha M beta 2 (CD11b/CD18, also called Mac-1), thus inhibiting mammalian neutrophil adhesion to endothelium (1). The A. caninum NIF cDNA encodes 257 amino acids (aa) including a 17 aa signal sequence and a 240 aa mature protein with 10 cysteine residues and 7 potential N-linked glycosylation sites (1). Incubation of mammalian neutrophils with NIF does not appear to be toxic, but does result in rapid, cation‑dependent, reversible inhibition of alpha M beta 2-mediated adhesion and degranulation (1 - 4). The NIF binding site within the alpha M beta 2 I-domain interferes with activation‑dependent adhesion sites of ligands including ICAM-1/CD54, complement component C3b, and fibrinogen (2, 3). In mouse or guinea pig inflammatory lung injury models, administration of NIF prevented neutrophil-dependent vascular injury (4, 5).

1. Moyle, M. et al. (1994) J. Biol. Chem. 269:10008.
2. Rieu, P. et al. (1994) J. Cell Biol. 127:2081.
3. Ustinov, V.A. and E.F. Plow (2002) J. Biol. Chem. 277:18769.
4. Barnard, J.W. et al. (1995) J. Immunol. 155:4876.
5. Zhou, M.-Y. et al. (1998) J. Clin. Invest. 101:2427.

Publications for NIF (5845-NF)(4)

Publications using 5845-NF

• Mackey-Alfonso, SE;Butler, MJ;Taylor, AM;Williams-Medina, AR;Muscat, SM;Fu, H;Barrientos, RM; Short-term high fat diet impairs memory, exacerbates the neuroimmune response, and evokes synaptic degradation via a complement-dependent mechanism in a mouse model of Alzheimer's disease Brain, behavior, and immunity 2024-07-21 [PMID: 39043341] (Bioassay, Mouse)
• Kirkland, JM;Edgar, EL;Patel, I;Kopec, AM; Impaired microglia-mediated synaptic pruning in the nucleus accumbens during adolescence results in persistent dysregulation of familiar, but not novel social interactions in sex-specific ways bioRxiv : the preprint server for biology 2023-05-03 [PMID: 37205324] (In Vivo, Rat)
• AM Kopec, CJ Smith, NR Ayre, SC Sweat, SD Bilbo Microglial dopamine receptor elimination defines sex-specific nucleus accumbens development and social behavior in adolescent rats Nat Commun, 2018-09-25;9(1):3769. 2018-09-25 [PMID: 30254300] (Cell Culture, In Vivo, Rat)
• D Wolf, N Anto-Miche, H Blankenbac, A Wiedemann, K Buscher, JD Hohmann, B Lim, M Bäuml, A Marki, M Mauler, D Duerschmie, Z Fan, H Winkels, D Sidler, P Diehl, DM Zajonc, I Hilgendorf, P Stachon, T Marchini, F Willecke, M Schell, B Sommer, C von Zur Mu, J Reinöhl, T Gerhardt, EF Plow, V Yakubenko, P Libby, C Bode, K Ley, K Peter, A Zirlik A ligand-specific blockade of the integrin Mac-1 selectively targets pathologic inflammation while maintaining protective host-defense Nat Commun, 2018-02-06;9(1):525. 2018-02-06 [PMID: 29410422] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • AAA27789(A. caninum)