PRODUCT CODE: EM1901-72

ROBO1 Monoclonal Antibody (EM1901-72)

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

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Western blot analysis of ROBO1 on Siha cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-72, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouset IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of ROBO1 on Siha cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-72, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouset IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of ROBO1 in SHSY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-72, 1/100) for 1 hour at room temperature, washed with PBS. Alexa FluorTM488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of ROBO1 in Siha cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-72, 1/100) for 1 hour at room temperature, washed with PBS. Alexa FluorTM488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human Liver cancer tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-72, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of ROBO1 was done on Siha cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-72, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa FluorTM488 Goat anti-Mouse IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of ROBO1 on Siha cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-72, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouset IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

ROBO1 Monoclonal Antibody (EM1901-72)

Immunogen

Recombinant protein within human robo1 aa 40-300.

Host

Mouse

Positive Control

Siha cell lysates, SHSY5Y, Siha, human Liver cancer tissue, human kidney tissue, human stomach cancer tissue, mouse brain tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

10E2

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein G purified.

MOLECULAR WEIGHT

Predicted band size 170-181 kDa.

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:1000-1:5000

  • IHC-P

  • 1:50-1:200

  • ICC

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

ROBO1

SYNONYMS

Deleted in U twenty twenty antibody; DUTT 1 antibody; DUTT1 antibody; FLJ21882 antibody; H Robo 1 antibody; H-Robo-1 antibody; hRobo 1 antibody; Robo 1 antibody; Robo1 antibody; ROBO1_HUMAN antibody; Roundabout 1 antibody; Roundabout axon guidance receptor homolog 1 antibody; Roundabout homolog 1 antibody; Roundabout homolog1 precurser antibody; Roundabout1 antibody; SAX 3 antibody; SAX3 antibody

SEQUENCE SIMILARITIES

Belongs to the immunoglobulin superfamily. ROBO family.

TISSUE SPECIFICITY

Widely expressed, with exception of kidney.

POST-TRANSLATIONAL MODIFICATION

Ubiquitinated. May be deubiquitinated by USP33.

SUBCELLULAR LOCATION

Membrane.

FUNCTION

Specific cells in the midline separate the left and right halves of the central nervous system and play many roles in guiding the growth cone behavior. In vertebrate spinal cord, insect abdominal nerves and nematodes, midline cells produce induced cues, such as nectins and slits, respectively, as attractants and repellents. respectively. These cells can serve as gatekeepers to prevent axons from passing through the midline and to guide the growth cone in response to the switching of lead cues beyond the crossing. One such gatherer, Robo, is an axon guidance receptor that defines a new subfamily of proteins from fruit flies to mammalian conserved Ig superfamily proteins. Robo acts as a receptor for the repellent Slit and functions in a cell-autonomous manner Non-intersecting axons express high levels of Robo and cross axons to express low levels of Robo and then reach midline and high levels of crossover. Robo1 and Robo2 are two human homologs of Drosophila megalopolis. Robo1 is also homologous to the C. elegans gene sax3, while Robo2 is homologous to zebrafish genes.