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) (SwissProt # Q9Y6N7). ROBO1 shows multiple isoform variants. The variant used here is termed ROBO1b/DUTT1 (Genbank # NP_598334), 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.
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Llama
Applications
Validated:
Western Blot, Immunocytochemistry
Cited:
Western Blot, Neutralization, Characterization
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ROBO1
Ser20-Pro861 (Val310-Ser312 del)
Accession # NP_598334
Ser20-Pro861 (Val310-Ser312 del)
Accession # NP_598334
Specificity
Detects human ROBO1 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant rat ROBO1 is observed, and less than 3% cross-reactivity with recombinant human (rh) ROBO2 and rhROBO3 is observed.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human ROBO1 Antibody
Detection of Human ROBO1 by Western Blot.
Western blot shows lysates of SH-SY5Y human neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7118) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ROBO1 at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.ROBO1 in HepG2 Human Cell Line.
ROBO1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Sheep Anti-Human ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7118) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.Applications for Human ROBO1 Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line
Western Blot
1 µg/mL
Sample: SH‑SY5Y human neuroblastoma cell line
Sample: SH‑SY5Y human neuroblastoma cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Sterile PBS to a final concentration of 0.2 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
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
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.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- 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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