PRODUCT CODE: em1901-72

ROBO1 Monoclonal Antibody (EM1901-72)

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

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

1000

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

Roundabout homolog 1

GENE NAME

ROBO1

SYNONYMS

Deleted in U twenty twenty,H-Robo-1,Roundabout homolog 1,ROBO1

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

Cell membrane; Single-pass type I membrane protein. Cell projection, axon. Endoplasmic reticulum-Golgi intermediate compartment membrane; Single-pass membrane protein. Note=Detected at growth cones in thalamus neurons. Detected at growth cones in thalamus neurons (By similarity). PRRG4 prevents cell surface location and both colocalize in the Endoplasmic reticulum/Golgi adjacent to the cell nucleus (By similarity).

FUNCTION

Receptor for SLIT1 and SLIT2 that mediates cellular responses to molecular guidance cues in cellular migration, including axonal navigation at the ventral midline of the neural tube and projection of axons to different regions during neuronal development. Interaction with the intracellular domain of FLRT3 mediates axon attraction towards cells expressing NTN1. In axon growth cones, the silencing of the attractive effect of NTN1 by SLIT2 may require the formation of a ROBO1-DCC complex (By similarity). Plays a role in the regulation of cell migration via its interaction with MYO9B; inhibits MYO9B-mediated stimulation of RHOA GTPase activity, and thereby leads to increased levels of active, GTP-bound RHOA. May be required for lung development (By similarity).