Skip to main content

Mouse ROBO3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF3155

R&D Systems, part of Bio-Techne
Catalog #
Availability
Size / Price
Qty
Loading...
AF3155
AF3155-SP

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Blocking, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived mouse ROBO3
Gly54-Ser545
Accession # Q9Z2I4

Specificity

Detects mouse ROBO3 in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross-reactivity with recombinant human (rh) ROBO3 and less than 5% cross-reactivity with rhROBO2 and rhROBO4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse ROBO3 Antibody

Detection of Mouse ROBO3 antibody by Western Blot.

Detection of Mouse ROBO3 by Western Blot.

Western blot shows lysates of Neuro-2A mouse neuroblastoma cell line. PVDF Membrane was probed with 0.5 µg/mL of Mouse ROBO3 Antigen Affinity-purified Polyclonal Antibody (Catalog # Af3155) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for ROBO3 at approximately 205 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
ROBO3 antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

ROBO3 in Mouse Embryo.

ROBO3 was detected in immersion fixed frozen sections of mouse embryo (E11.5) using Mouse ROBO3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3155) at 25 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to developing forebrain. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Applications for Mouse ROBO3 Antibody

Application
Recommended Usage

Immunohistochemistry

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

Western Blot

0.5 µg/mL
Sample: Neuro‑2A mouse neuroblastoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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.

Reconstitution Buffer Available:
Size / Price
Qty
Loading...

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.

Background: ROBO3

Mouse ROBO3 (also named Rig-1) 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). Mouse ROBO3 is a type I transmembrane glycoprotein that is synthesized as a 1366 amino acid (aa) precursor. It contains a 20 aa signal sequence, an 871 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 454 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 ROBO-1 (7, 8). Mouse ROBO3 has multiple isoforms. An alternate start site generates an 1366 aa (precursor) A isoform and a 1344 aa (precursor) B isoform. These two forms only differ in the first 53 and 31 amino acids of the precursor, respectively (9). There are reportedly nine splice variants in the mouse ROBO3 gene. Three result in soluble forms. Little information exists about the isoforms. Mouse ROBO3 ECD is 95%, 84% and 86% aa identical to the ROBO3 ECD in rat, human and canine, 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 ROBO-1, blocking SLIT activation. A second possibility involves ROBO3 binding to SLIT2 in a nonproductive interaction. In human, 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. Yuan, S-S.F. et al. (1999) Dev. Biol. 207:62.
  6. SwissProt. Accession # Q9Z2I4.
  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

Additional ROBO3 Products

Product Documents for Mouse ROBO3 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse ROBO3 Antibody

For research use only

Loading...
Loading...
Loading...
Loading...