Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 356001
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Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human ROBO2 isoform 1
Ser22-Pro859
Accession # Q9HCK4.2

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human ROBO2 Antibody

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

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.

Applications for Human ROBO2 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 human cell line transfected with human ROBO2 and eGFP

Western Blot

1 µg/mL
Sample: Recombinant Human ROBO2 Fc Chimera (Catalog # 3147-RB)

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

SAX3

Entrez Gene IDs

6092 (Human); 268902 (Mouse); 84409 (Rat)

Gene Symbol

ROBO2

UniProt

Additional ROBO2 Products

Product Documents for Human ROBO2 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human ROBO2 Antibody

For research use only

Citations for Human ROBO2 Antibody

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