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Human ROBO1 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF7118

R&D Systems, part of Bio-Techne
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AF7118
AF7118-SP

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Llama

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Characterization, Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human ROBO1
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 antibody by Western Blot.

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 antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC).

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

Western Blot

1 µg/mL
Sample: SH‑SY5Y human neuroblastoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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.

Reconstitution Buffer Available:
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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.

Background: ROBO1

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.

Long Name

Roundabout Homolog 1

Alternate Names

DUTT1

Entrez Gene IDs

6091 (Human); 58946 (Rat)

Gene Symbol

ROBO1

Additional ROBO1 Products

Product Documents for Human ROBO1 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 Human ROBO1 Antibody

For research use only

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