Recombinant Human Slit3 (aa 34-1116) His-tag Protein, CF

Recombinant Human Slit3 His-tag Protein (Catalog # 9255-SLB) Induces Cortical Neurite Outgrowth. A) Neurite outgrowth in E16-18 embryonic rat cortical neurons treated with Recombinant Human Slit3 His-tag Protein. B) Untreated E16-18 embryonic rat cortical neurons.

2 μg/lane of Recombinant Human Slit3 His-tag Protein (Catalog # 9255-SLB) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 119-149 kDa.

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

Recombinant Human Slit3 (aa 34-1116) His-tag Protein, CF Summary

• Details of Functionality: Measured by its ability to enhance neurite outgrowth of E16-E18 rat embryonic cortical neurons. Recombinant Human Slit3 His-tag (Catalog # 9255-SLB) enhances neurite outgrowth at 5 μg/mL.
• Source: Human embryonic kidney cell, HEK293-derived human Slit3 protein
  Cys34-Val1116, with a C-terminal 6-His tag
• Accession #: NP_003053.1
• N-terminal Sequence: Cys34
• 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: 121 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: 119-149 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 and NaCl 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 Slit3 (aa 34-1116) His-tag Protein, CF

• KIAA0814
• MEGF5
• MEGF5Multiple epidermal growth factor-like domains protein 5
• Multiple EGF-like domains protein 5
• SLIL2
• SLIL2FLJ10764
• slit homolog 3 (Drosophila)
• slit homolog 3 protein
• SLIT1
• slit2
• Slit3
• Slit-3slit (Drosophila) homolog 3

Background

Slit3 is an approximately 200 kDa member of the Slit family of large secreted axon guidance molecules that are ligands for ROBO receptors (1, 2). It contains a mitochondrial targeting sequence and may also be retained intracellularly (3). Mature human Slit3 consists of 4 cassettes of leucine-rich repeats (LRR) flanked by LRR N-terminal and C-terminal domains, followed by multiple EGF-like domains, a Laminin G-like domain, and a C-terminal cysteine-rich domain. Alternative splicing generates additional isoforms with a substituted cysteine-rich domain or a deletion in the last EGF-like domain and Laminin G-like domain. During development, Slit3 is expressed in the ventral neural tube, developing sensory organs, limb buds, and developing areas of the limbs in patterns that overlap with but are discrete from Slit1 and Slit2 (1, 2, 4). Axons will not be allowed to recross the floor plate unless all three Slit genes are disrupted, suggesting some overlap in Slit function (5). Slit3 is also expressed in the lung, kidney, skeletal muscle, and heart, both during development and postnatally (6-8). Mice with genetically disrupted Slit3 show abnormalities in diaphragm and kidney development (7, 8). Slit3 is additionally expressed by vascular endothelial cells and smooth muscle cells (9). It binds to both ROBO1 and ROBO4, but it is the ROBO4 interaction that mediates Slit3-induced angiogenesis (9). It also can enhance the chemokine-induced migration of monocytes (10). The related Slit2 protein is cleaved in vivo (at a site conserved in Slit3), and the resulting C-terminal fragment binds Plexin A1 and retains the ability to repel axon migration (4, 11). The corresponding C-terminal fragment of Slit3 is able to bind heparin and neutralize its anti-coagulant activity (12). Within this fragment
(aa 1120-1523), human Slit3 shares 93% amino acid identity with mouse and rat Slit3.

1. Blockus, H. and A. Chedotal (2014) Curr. Opin Neurobiol. 27:82.
2. Gara, R.K. et al. (2015) Drug Discov. Today 20:156.
3. Little, M. H. et al. (2001) Am. J. Physiol. Cell Physiol. 281:C486.
4. Brose, K. et al. (1999) Cell 96:795.
5. Long, H. et al. (2004) Neuron 42:213.
6. Greenberg, J.M. et al. (2004) Dev. Dyn. 230:350.
7. Liu, J. et al. (2003) Mech. Dev. 120:1059.
8. Yuan, W. et al. (2003) Proc. Natl. Acad. Sci. USA 100:5217.
9. Zhang, B. et al. (2009) Blood 114:4300.
10. Geutskens, S.B. et al. (2010) J. Immunol. 185:7691.
11. Delloye-Bourgeois, C. et al. (2015) Nat. Neurosci. 18:36.
12. Condac, E. et al. (2012) Glycobiology 22:1183.

Bioinformatics

• Uniprot:
  • NP_003053.1()