Anti-Ribonuclease 3 antibody [5G4]
cat.: EM1801-17
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human
Applications: WB, ICC, IHC-P, FC
Clonality: Monoclonal
Clone number: 5G4
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 purified.
Molecular weight: 18 kDa
Isotype: IgG1
Immunogen: Recombinant protein within human Ribonuclease 3 aa 28-120.
Positive control: U937, Hela, SHG-44, human spleen tissue, human kidney tissue.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  ICC
  IHC-P
  FC

1:500-1:2000
1:100
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P12724 Human
Alternative names: Cytotoxic ribonuclease antibody ECP antibody ECP_HUMAN antibody Eosinophil cationic protein antibody OTTHUMP00000164017 antibody Ribonuclease 3 antibody Ribonuclease, RNase A family, 3 antibody RNase 3 antibody RNASE3 antibody RNS3 antibody
Images
EM1801-17_1.jpg Fig1: Western blot analysis of Ribonuclease 3 on U937 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 was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Lane 1: Anti-Ribonuclease 3 Antibody, (1:500).
Lane 2: Anti-Ribonuclease 3 Antibody, (1:500), preincubated with the immunization protein.
EM1801-17_2.jpg Fig2: ICC staining Ribonuclease 3 in Hela 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 Ribonuclease 3 monoclonal antibody at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
EM1801-17_3.jpg Fig3: ICC staining Ribonuclease 3 in SHG-44 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 Ribonuclease 3 monoclonal antibody at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
EM1801-17_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Ribonuclease 3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 antibody (EM1801-17) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
EM1801-17_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Ribonuclease 3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 antibody (EM1801-17) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
EM1801-17_6.jpg Fig6: Flow cytometric analysis of Ribonuclease 3 was done on Hela cells. The cells were fixed, permeabilized and stained with Ribonuclease 3 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-Mouse IgG Secondary antibody at 1/500 dilution for 30 minutes.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.