Human ROBO3 Biotinylated Antibody

Catalog #: BAF3076 Datasheet / COA / SDS

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BAF3076

All ROBO3 Antibodies

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Human ROBO3 Biotinylated Antibody Summary

Species Reactivity

Human

Specificity

Detects human ROBO3 in Western blots. In Western blots, approximately 60% cross‑reactivity with recombinant mouse ROBO3 is observed, 10% cross-reactivity with recombinant rat ROBO1 is observed, and less than 1% cross-reactivity with recombinant human (rh) ROBO2 and rhROBO4 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

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

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Label

Biotin

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human ROBO3 Fc Chimera (Catalog # 3076-RB)

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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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

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).

Long Name

Roundabout Homolog 3

Entrez Gene IDs

64221 (Human); 19649 (Mouse)

Alternate Names

FLJ21044; HGPPS; HGPS; RBIG1horizontal gaze palsy with progressive scoliosis; retinoblastoma inhibiting gene 1; RIG1; ROBO3; roundabout (axon guidance receptor, Drosophila) homolog 3; roundabout homolog 3; roundabout, axon guidance receptor, homolog 3 (Drosophila); Roundabout-like protein 3