ROBO1 Mouse Monoclonal Antibody [10E2]
cat.: EM1901-72
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IHC-P, ICC, FC
Clonality: Monoclonal
Clone number: 10E2
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2 mg/ml.
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 181 kDa
Isotype: IgG1
Immunogen: Recombinant protein within Human ROBO1 aa 41-286 / 1,651.
Positive control: 293T cell lysate, Hela cell lysate, PC-3 cell lysate, SHSY5Y, SiHa, human Liver cancer tissue, human kidney tissue, human stomach cancer tissue, mouse brain tissue, Siha.
Subcellular location: Cell membrane, Cell projection, axon, Endoplasmic reticulum-Golgi intermediate compartment membrane.
Recommended Dilutions:
  WB
  IHC-P
  ICC
  FC

1:1000-1:5000
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: Q9Y6N7 Human | O89026 Mouse
Alternative names: Deleted in U twenty twenty DUTT 1 DUTT1 FLJ21882 H Robo 1 H-Robo-1 hRobo 1 Robo 1 Robo1 ROBO1_HUMAN Roundabout 1 Roundabout axon guidance receptor homolog 1 Roundabout homolog 1 Roundabout homolog1 precurser Roundabout1 SAX 3 SAX3
Images
EM1901-72_1.jpg Fig1: Western blot analysis of ROBO1 on different lysates with Rabbit anti-ROBO1 antibody (EM1901-72) at 1/500 dilution.

Lane 1: 293T cell lysate
Lane 2: Hela cell lysate
Lane 3: PC-3 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 181 kDa
Observed band size: 181 kDa

Exposure time: 30 seconds;

6% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-72) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
EM1901-72_2.jpg Fig2: 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).
EM1901-72_3.jpg Fig3: 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).
EM1901-72_4.jpg Fig4: 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.
EM1901-72_5.jpg Fig5: 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.
EM1901-72_6.jpg Fig6: 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.
EM1901-72_7.jpg Fig7: 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.
EM1901-72_8.jpg Fig8: 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).
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