Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.20 EU per 1 μg of the antibody by the LAL method.
Measured by its ability to neutralize the the enhancemment of neurite outgrowth of dorsal root ganglion neurons from E13 chick embryos induced by Slit3. The Neutralization Dose (ND50) is typically 25 µg/mL in the presence of 25 µg/mL Recombinant Mouse Slit3.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Rat Slit3 by Western Blot. Western blot shows lysates of rat embryonic cortical glial cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Slit3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3629) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for Slit3 at approximately 100 kDa and 160 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
Slit3 is a member of the slit family of large secreted axon guidance molecules that are ligands for Robo receptors (1). Like other mammalian family members, the 1523 amino acid (aa) Slit3 contains a signal sequence followed by 23 leucine-rich repeats (LRR) and nine EGF-like sequences (1). Mammalian Slits also contain a laminin-G domain between EGF6 and EGF7, and a C-terminal cysteine-rich domain (1). In Drosophila Slit, specific LRR are sites of ROBO interaction and homodimerization (2). Drosophila Slit shows equal similarities with all three mouse slit proteins, which are 59‑66% identical with each other (1). 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). Axons will not be allowed to recross the floor plate unless all three Slit genes are disrupted, suggesting some overlap in function (3). Slit3 is also expressed in the lung, kidney, skeletal muscle and heart, both during development and postnatally (1, 4‑6). ROBO2 is often found in complementary areas (4). Mice with genetically disrupted Slit3 have thin diaphragms with disorganized collagen fibrils and frequently develop diaphragmatic hernias (5, 6). Abnormalities in kidney development may also occur (5). Although Slit proteins are generally considered to be secreted, significant amounts of Slit3 may be retained in the mitochondria because of features in the signal sequence that indicate a high probability of mitochondrial targeting (7). Secreted Slit is often membrane-associated (7). Mouse Slit3 shows 98% and 94% amino acid identity with rat and human Slit3, respectively.
Yuan, W. et al. (1999) Dev. Biol. 212:290.
Howitt, J. A. et al. (2004) EMBO J. 23:4406.
Long, H. et al. (2004) Neuron 42:213.
Greenberg, J. M. et al. (2004) Dev. Dyn. 230:350.
Liu, J. et al. (2003) Mech. Dev. 120:1059.
Yuan, W. et al. (2003) Proc. Natl. Acad. Sci. USA 100:5217.
Little, M. H. et al. (2001) Am. J. Physiol. Cell Physiol. 281:C486.
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