Recombinant Human MuSK Fc Chimera Avi-tag Protein, CF

When Biotinylated Recombinant Human MuSK Fc Chimera Avi-tag Protein (Catalog # AVI9810) is immobilized onto a Streptavidin coated plate (CP004) at 2 µg/mL (100 µL/well), in the presence of Recombinant Human Agrin (6624-AG), it binds to Recombinant Human LRP-4 (5948-LR) with an ED50 of 0.600-4.80 ng/mL.

2 μg/lane of Biotinylated Recombinant Human MuSK Fc Chimera Avi-tag Protein (Catalog # AVI9810) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 87-102 kDa and 170-200 kDa, respectively.

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

Recombinant Human MuSK Fc Chimera Avi-tag Protein, CF Summary

• Additional Information: Biotinylated
• Details of Functionality: Measured by its binding ability in a functional ELISA. When Biotinylated Recombinant Human MuSK Fc Chimera Avi-tag (Catalog # AVI9810) is immobilized onto a Streptavidin coated plate (Catalog # CP004) at 2 µg/mL (100 µL/well), in the presence of Recombinant Human Agrin (Catalog # 6624-AG), it binds to Recombinant Human LRP-4 (Catalog # 5948-LR) with an ED₅₀ of 0.600-4.80 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human MuSK protein
  

  Human MuSK (Leu24-Thr495) Accession # O15146.1	IEGRMD	Human IgG1 (Pro100-Lys330)	Avi-tag
  N-terminus			C-terminus

• N-terminal Sequence: Leu24
• Structure / Form: Disulfide-linked homodimer
  Biotinylated via Avi-tag
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 80 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: 87-102 kDa, under 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 with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Reconstitution Instructions: Reconstitute at 500 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 Human MuSK Fc Chimera Avi-tag Protein, CF

• EC 2.7.10
• EC 2.7.10.1
• MGC126323
• MGC126324
• muscle, skeletal, receptor tyrosine kinase
• MuSK
• skeletal receptor tyrosine-protein kinase

Background

MuSK (muscle-specific kinase) is a 100 kDa type I transmembrane (TM) protein belonging to the receptor tyrosine kinase family (1). It contains a 472 aa extracellular domain (ECD), a 21 aa transmembrane domain, and a 353 cytoplasmic domain (1). Within the ECD, Human MuSK shares 91% and 92% aa sequence identity with mouse MuSK and rat MuSK, respectively. It is found in the postsynaptic membrane of skeletal muscle motor endplates (2). Human MuSK has multiple isoforms. One contains deletions at residues 307‑394 and 454‑461, while a second is a short soluble form that contains residues 120‑209 plus a unique 24 aa C‑terminal tail (3). MuSK binds the heparin sulfate proteoglycan agrin to promote acetylcholine receptor clustering. It has also been found to bind with low-density lipoprotein receptor-related protein 4 (LRP4) (4), Wnt ligands (5), biglycan (6), and ColQ (7). Recent studies have showed that MuSK does not bind agrin directly, but enhanced the MuSK-LRP4 interaction (7, 8). When Agrin binds to the N-Terminal region of LRP4, this promotes the association of LRP4 and MuSK, which then stimulates MuSK kinase activity (9, 10). Our Avi-tag Biotinylated human MUSK Fc Chimera features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

1. Valenzuela, D.M. et al. (1995) Neuron. 15:573.
2. Finn, A.J. et al. (2003) Nat. Neurosci. 6:717.
3. Nasrin, F. et al. (2014) Sci. Rep. 4:6841.
4. Kim, N. et al. (2008) Cell 135:334.
5. Banerjee, S. et al. (2011) Development 138:3287.
6. Amenta, A.R. et al. (2012) J Neurosci 32:2324.
7. Otsuka, K. et al (2015) Scientific Reports. 5:13928.
8. Zhang, B. et al. (2008) Neuron 60:285.
9. Zhang, W. et al. (2011) J Biol Chem. 286(47):40624.
10. Burden, S.J. et al. (2017) CSH Perspectives Biol. 5:a009167.