Human ROBO3 Antibody

R&D Systems | Catalog # AF3076

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Avian - Chicken, Chicken, Drosophila, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all ROBO3 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human ROBO3
Leu40-Ser545
Accession # Q96MS0

Specificity

Detects ROBO3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 60% cross-reactivity with recombinant mouse ROBO3 is observed, 5% cross-reactivity with recombinant human ROBO2 and recombinant rat ROBO1 is observed, and less than 1% cross‑reactivity with recombinant human ROBO4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human ROBO3 Antibody

Detection of ROBO3 by Immunohistochemistry

Detection of ROBO3 by Immunohistochemistry

Histology and labeling of axon tracts in the brain of control and Ftm−/− fetuses. A–D, Nissl staining on coronal sections of the brain at two distinct anteroposterior levels of thalamic and hypothalamic regions in E18.5 WT (A, C) and Ftm−/− (B, D) fetuses. C, D, More posterior sections than A and B. Both levels of sections correspond to the ventral hypothalamus and the alar thalamus. Black arrowheads in B–D point to axon fascicles of the IC and RT. Double black arrows in B and D point to the dysmorphic hypothalamus in Ftm mutants. E–F′, Carbocyanine dye staining of corticothalamic (DiI, magenta) and thalamocortical (DiA, green) axons in E18.5 WT (E, E′) and Ftm−/− (F, F′) brains. E′, F′, Higher-magnification of the boxed regions in E and F, respectively. G, H, Neurofilament (NF) immunostaining of axon tracts in E18.5 control (G) and Ftm−/− (H) brains. I–L′, Immunofluorescence for Tuj1 and NF (I–J′) and for Robo3 and Tag1 (K–L′) in E13.5 control (K, K′) and Ftm−/− (L, L′) brains. M–N′, Nissl staining on coronal sections at the level of the eyes of the head of E18.5 WT (M, M′) and Ftm−/− (N, N′) fetuses. M′, N′, Higher-magnification of the boxed regions in M and N, respectively. The arrowhead in N′ points to remnants of the RPE. AN, Anteroventral nucleus; CX, cortex; HB, Habenula; NR, neural retina; V3 third ventricle. Scale bars: (in A, M) A–D, M, N, 1 mm; (in E, G) E–H, 0.5 mm; (in E′, I, K′), E′, F′, I–J′, K′, L′, 0.1 mm; (in K, M′) K, L, M′, N′, 0.2 mm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30692221), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ROBO3 by Immunohistochemistry

Detection of ROBO3 by Immunohistochemistry

Medial habenular volumes analysis in Wnt1–/–. Lateral views of E18.5 wt (A) and Wnt1–/–(B) brains analyzing ROBO3 distribution by iDISCO. We can observe the mHB and the initial fr portion. (C) Significance among wt (n = 6) and Wnt1–/– mHb volumes (n = 4) was analyzed by an unpaired t-test. Significant differences were found (*⁣*⁣**< 0.0001), being the mutant mHb volume larger than the wt. fr, fasciculus retroflexus; mHb, medial habenula. Scale bar: 200 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34722541), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human ROBO3 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed frozen sections of embryonic mouse spinal cord (E11.5)

Western Blot

0.1 µg/mL
Sample: Recombinant Human ROBO3 Fc Chimera (Catalog # 3076-RB)

Reviewed Applications

Read 1 review rated 5 using AF3076 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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: ROBO3

Human ROBO3 is a 200 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 ROBO3 is a type I transmembrane glycoprotein that is synthesized as a 1386 amino acid (aa) precursor. It contains a 20 aa signal sequence, an 871 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 474 aa cytoplasmic region (5, 6). The ECD contains five C2-type Ig-like domains (aa 64‑531) and three fibronectin (FN) type III domains (aa 555‑863). The cytoplasmic region contains three of four possible 15‑20 aa long CC (conserved cytoplasmic) motifs that are found in ROBO1 (7, 8). Human ROBO3 has multiple isoforms. An alternate start site generates a 1365 aa A isoform and a 1341 aa B isoform. These two mature forms differ only over the first 26 and 2 amino acids of the N-terminus, respectively. There are multiple point mutations and insertions in the ROBO3 gene. Three result in truncated forms. One is 456 aa in length and ends after the Ig-like domain # 4. A second is 770 aa in length and ends in the third FN domain. A third isoform is truncated after aa 1108 in the cytoplasmic region after CC2. At least one alternate splice form is also reported. It shows a 10 aa substitution between aa 1025‑1034, followed by truncation. Human ROBO3 ECD is 84% and 91% aa identical to the ECD in mouse and canine ROBO3, respectively. Normally, axons originating on one side of the spinal cord are inhibited from crossing to the other side by a SLIT2-ROBO1 interaction at the midline. ROBO3 is permissive for this event. It is unclear how this is accomplished. One possibility is that it binds directly to ROBO1, blocking SLIT activation. A second possibility involves ROBO3 binding to SLIT2 in a nonproductive interaction. However, only ROBO3 Form B is known to bind to SLIT2 (9‑11).

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. Jen, J.C. et al. (2004) Science 304:1509.
  6. Swiss-Prot. Accession # Q96MS0.
  7. Bashaw, G.J. et al. (2000) Cell 101:703.
  8. Kidd, T. et al. (1998) Cell 92:205.
  9. Camurri, L. et al. (2005) Mol. Cell. Neurosci. 30:485.
  10. Sabatier, C. et al. (2004) Cell 117:157.
  11. Mambetisaeva, E.T. et al. (2005) Dev. Dyn. 233:41.

Long Name

Roundabout Homolog 3

Alternate Names

RIG1

Entrez Gene IDs

64221 (Human); 19649 (Mouse)

Gene Symbol

ROBO3

UniProt

Q96MS0

Additional ROBO3 Products

Product Documents for Human ROBO3 Antibody

Certificate of Analysis

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Product Specific Notices for Human ROBO3 Antibody

For research use only

Related Research Areas

Neuronal Lineage Markers

Citations for Human ROBO3 Antibody

Customer Reviews for Human ROBO3 Antibody (1)

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  • Human ROBO3 Antibody
    Name: Ricardo Vieira
    Application: Immunohistochemistry-Frozen
    Sample Tested: Embryonic spinal cord
    Species: Mouse
    Verified Customer | Posted 05/08/2019
    E11.5 Mouse spinal cord. In green, Comissural neurons are labelled using the antibody against Robo-3.
    Human ROBO3 Antibody AF3076

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