ROBO1 (Roundabout-like protein 1) is a 190-230 kDa member of the ROBO/roundabout receptor family. It is expressed by commissural axons from multiple nuclei, and is also found on vascular endothelium, bronchial epithelium, and synctiotrophoblasts. It interacts with Slit and DCC to temporally regulate the migration of axonal processes. The human ROBO1 precursor is a 1651 amino acid (aa) type I transmembrane protein. It contains a 25 aa signal sequence, followed by an 872 aa extracellular region (aa 26-897) that possesses five C2-type Ig-like domains (aa 68-541) and three fibronectin type III domains (aa 561-864). ROBO1 shows multiple isoform variants. The variant used here is termed ROBO1b/DUTT1, and it possesses an 18 aa substitution for aa 1‑47, accompanied by a three aa insertion after Gln348, and a deletion of aa 939-947. ROBO1a, by contrast, possesses only the 18 aa substitution just described. A third isoform possesses the same changes as ROBO1b plus an additional deletion of aa 1013-1067, while a final variant utilizes an alternative start site at Met120. Proteolytic cleavage generates a soluble 120 kDa N-terminal fragment. Over aa 20-861, human ROBO1b shares 97% aa identity with mouse ROBO1.
Human ROBO1 Antibody
R&D Systems | Catalog # MAB7118
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Western Blot
Cited:
Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 770506
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ROBO1b
Ser20-Pro861 (Val310Ser312 del)
Accession # NP_598334
Ser20-Pro861 (Val310Ser312 del)
Accession # NP_598334
Specificity
Detects human ROBO1 in direct ELISAs.
In direct ELISAs, approximately 50% cross-reactivity
with recombinant rat ROBO1 is observed and no cross-reactivity with recombinant
human (rh) ROBO2 or rhROBO3 is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Scientific Data Images for Human ROBO1 Antibody
Detection of Human ROBO1 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and SH-SY5Y human neuroblastoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ROBO1 Monoclonal Antibody (Catalog # MAB7118) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for ROBO1 at approximately 220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Detection of Human ROBO1 by Western Blot
NRP1, ROBO1, and ITG alpha 5 were identified as target genes of miR-148a/b-3p.a Venn diagrams illustrate the intersection between three algorithms that were used to predict potential gene targets of miR-148a/b-3p. b RT-PCR analysis indicates that the expression of NRP1, ROBO1, and ITG alpha 5 increased during UCB-MNC to OEC differentiation. c The mRNA levels of NRP1, ROBO1, and ITGɑ5 in ECs were regulated by miR-148a/b. ECs were transfected with miR mimics (80 µM), and 48 h after transfection, endogenous NRP1 expression was detected by RT-PCR. d At 24, 48, or 72 h after transfection, Western blotting analysis was performed to investigate the protein levels of target genes. The internal loading control was beta -actin. e Relative mRNA expression of NRP1, ROBO1, and ITGɑ5 was confirmed by RT-PCR. PECAM1 expression was unchanged following treatment with the miR-148a/b-3p mimic (n = 5 per group). f Quantification of the luciferase reporter assay data comparing the wildtype (Wt) NRP1-3′UTR and a mutated (Mut) NRP1-3′UTR; the (Mut) contained a 7-nucleotide mutation in the predicted miR-148a/b-3p binding site. The schematic depicts the predicted miR-148a/b-3p binding site (blue) in the NRP1-3′UTR and shows the 7-nucleotide mutation. All data are presented as the mean ± SEM, **P < 0.01, ***P < 0.001 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31723119), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human ROBO1 Antibody
Application
Recommended Usage
Western Blot
2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and SH‑SY5Y human neuroblastoma cell line
Sample: HeLa human cervical epithelial carcinoma cell line and SH‑SY5Y human neuroblastoma cell line
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL. 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
Background: ROBO1
Long Name
Roundabout Homolog 1
Alternate Names
DUTT1
Gene Symbol
ROBO1
UniProt
Additional ROBO1 Products
Product Documents for Human ROBO1 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human ROBO1 Antibody
For research use only
Citations for Human ROBO1 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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