ROBO1 Antibody (770506)

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.

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 (//pubmed.ncbi.nlm.nih.gov/31723119), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB
• Clone: 770506
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

ROBO1 Antibody (770506) Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human ROBO1b
  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.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: ROBO1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ROBO1 Antibody (770506)

• DUTT1
• DUTT1Drosophila) homolog 1
• H-Robo-1
• MGC131599
• MGC133277
• ROBO1
• roundabout, axon guidance receptor, homolog 1 (Drosophila)

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ROBO1 Antibody (MAB7118)(4)

Publications using MAB7118

• H Kim, Y Ko, H Park, H Zhang, Y Jeong, Y Kim, M Noh, S Park, YM Kim, YG Kwon MicroRNA-148a/b-3p regulates angiogenesis by targeting neuropilin-1 in endothelial cells Exp. Mol. Med., 2019-11-13;51(11):134. 2019-11-13 [PMID: 31723119] (Western Blot, Human)
• G Genet, K Boyé, T Mathivet, R Ola, F Zhang, A Dubrac, J Li, N Genet, L Henrique G, L Benedetti, S Künzel, L Pibouin-Fr, JL Thomas, A Eichmann Endophilin-A2 dependent VEGFR2 endocytosis promotes sprouting angiogenesis Nat Commun, 2019-05-28;10(1):2350. 2019-05-28 [PMID: 31138815] (Western Blot, Human)
• Feng Xu, Ling-Yun Wu, Chun-Kang Chang, Qi He, Zheng Zhang, Li Liu et al. Whole-exome and targeted sequencing identify ROBO1 and ROBO2 mutations as progression-related drivers in myelodysplastic syndromes Nature Communications 11/26/2015 [PMID: 26608094]
• Rama N, Dubrac A, Mathivet T, Ni Charthaigh R, Genet G, Cristofaro B, Pibouin-Fragner L, Ma L, Eichmann A, Chedotal A Slit2 signaling through Robo1 and Robo2 is required for retinal neovascularization. Nat Med, 2015-04-20;21(5):483-91. 2015-04-20 [PMID: 25894826] (Western Blot, Human)

Bioinformatics

• Gene Symbol: ROBO1
• Uniprot:
  • NP_598334(Human)