Human ROBO2 Antibody

Catalog # Availability Size / Price Qty
MAB3147
MAB3147-SP
Detection of ROBO2 in HEK293 Human Cell Line transfected with Human ROBO2 and eGFP by Flow Cytometry.
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Product Details
Citations (3)
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Human ROBO2 Antibody Summary

Species Reactivity
Human
Specificity
Detects human ROBO2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) ROBO1, rhROBO3, or rhROBO4 is observed.
Source
Monoclonal Mouse IgG1 Clone # 356001
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ROBO2 isoform 1
Ser22-Pro859
Accession # Q9HCK4.2
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
Recombinant Human ROBO2 Fc Chimera (Catalog # 3147-RB)
Flow Cytometry
0.25 µg/106 cells
HEK293 human cell line transfected with human ROBO2 and eGFP
Immunohistochemistry
8-25 µg/mL
Immersion fixed paraffin-embedded sections of human brain (cortex)
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
8-25 µg/mL
Immersion fixed rat cortical stem cells differentiated for 7‑14 days by growth factor withdrawal

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.

Scientific Data

Flow Cytometry Detection of ROBO2 antibody in HEK293 Human Cell Line transfected with Human ROBO2 and eGFP antibody by Flow Cytometry. View Larger

Detection of ROBO2 in HEK293 Human Cell Line transfected with Human ROBO2 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human ROBO2 or (B) irrelevant protein, and eGFP, was stained with Mouse anti-Human ROBO2 monoclonal antibody (Catalog # MAB3147) followed by Allophycocyanin-conjugated anti-Mouse IgG secondary antibody (F0101B). Quadrant markers were set based on isotype control antibody staining (MAB002, data not shown). Staining was performed using our Staining Membrane-Associated Proteins protocol.

Immunohistochemistry View Larger

ROBO2 in Human Brain (Cortex). ROBO2 was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Mouse Anti-Human ROBO2 Monoclonal Antibody (Catalog # MAB3147) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). 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 cytoplasm in neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
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.

Background: ROBO2

Human ROBO2 is a 175 kDa member of the ROBO family of guidance molecules (1‑3). The term ROBO derives from round-about, a description of the circuitous pathway axons take in the absence of a functional ROBO gene (3, 4). Human ROBO2 is a type I transmembrane (TM) glycoptotein that is synthesized as a 1378 amino acid (aa) precursor. It contains a 21 aa signal sequence, an 838 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 498 aa cytoplasmic region (5, 6). The ECD contains five C2-type Ig-like domains (aa 31‑504) and three fibronectin (FN) type III domains (aa 522‑826). The cytoplasmic region contains multiple 15‑20 aa long CC (conserved cytoplasmic) motifs (C0‑C2) that are found in ROBO-1 (7, 8). Human ROBO2 has at least two potential isoforms. One isoform shows a cytoplasmic truncation of the C-terminus (aa 1186‑1378) (9). A second isoform is a 570 aa soluble form that shows a deletion of the first two and one-half C2-type Ig-like domains (aa 22‑285) and terminates after the third fibronectin domain with a unique 20 aa sequence (10). Based on mouse and rat ROBO2 GenBank sequences, considerably more alternate splice forms may occur. Human ROBO2 ECD shares 98% aa sequence identity with the ECD in mouse and canine ROBO2. ROBO2 would appear to play a number of roles in cell adhesion. In the neural tube, it is the receptor for a chemorepulsant. Axons that cross the midline are directed, presumably by ROBO2/SLIT chemorepulsion, to lateral positions in the contralateral spinal cord (6, 11, 12). In addition, ROBO2 is both permissive for neurite outgrowth (via ROBO1‑ROBO2 interaction) and inhibitory for neurite outgrowth (via ROBO2-SLIT interaction) (13, 14).

References
  1. Rajagopalan, S. et al. (2000) Neuron 28:767.
  2. Guthrie, S. (2004) Curr. Biol. 14:R632.
  3. Guthrie, S. (2001) Curr. Biol. 11:R300.
  4. Seeger, M. et al. (1993) Neuron 10:409.
  5. Kidd, T. et al. (1998) Cell 92:205.
  6. Nguyen Ba-Charvet, K.T. et al. (2001) J. Neurosci. 21:4281.
  7. Bashaw, G.J. et al. (2000) Cell 101:703.
  8. Jen, J.C. et al. (2004) Science 304:1509.
  9. GenBank Accession # Q9KCK4.
  10. Isogai, T. et al. (2002) GenBank Accession # AK074780.
  11. Mambetisaeva, E.T. et al. (2005) Dev. Dyn. 233:41.
  12. Long, H. et al. (2004) Neuron 42:213.
  13. Hivert, B. et al. (2002) Mol. Cell. Neurosci. 21:534.
  14. Lin, L. et al. (2005) Mol. Cell. Neurosci. 28:547.
Long Name
Roundabout Homolog 2
Entrez Gene IDs
6092 (Human); 268902 (Mouse); 84409 (Rat)
Alternate Names
Drosophila) homolog 2; ROBO2; roundabout, axon guidance receptor, homolog 2 (Drosophila); SAX3

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Citations for Human ROBO2 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. SLIT2/ROBO2 signaling pathway inhibits nonmuscle myosin IIA activity and destabilizes kidney podocyte adhesion
    JCI Insight, 2016-11-17;1(19):e86934.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  2. COUP-TFII regulates satellite cell function and muscular dystrophy
    J Clin Invest, 2016-09-12;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: IHC
  3. The axonal repellent Slit2 inhibits pericyte migration: potential implications in angiogenesis.
    Authors: Guijarro-Munoz I, Cuesta AM, Alvarez-Cienfuegos A, Geng JG, Alvarez-Vallina L, Sanz L
    Exp. Cell Res., 2011-12-14;318(4):371-8.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

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