Application
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.Application
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).Application
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).Application
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.Application
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.Application
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.| Product Name | Ribonuclease 3 [5G4] |
|---|---|
| Antibody Type | Primary Antibodies |
| Antigen Alias | 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 |
| Product description | The protein encoded by this gene belongs to the pancreatic ribonuclease family, a subset of the ribonuclease A superfamily. Cytotoxin and helminthotoxin with low-efficiency ribonuclease activity. The protein exhibits antimicrobial activity against pathogenic bacteria. Exhibits antibacterial activity, including cytoplasmic membrane depolarization of preferentially Gram-negative, but also Gram-positive strains. Promotes E.coli outer membrane detachment, alteration of the overall cell shape and partial loss of cell content. ECP is a potent cytotoxic protein capable of killing cells of guinea pig tracheal epithelium, mammalian leukemia, epidermis carcinoma, and breast carcinoma, as well as non-mammalian cells such as parasites, bacteria, and viruses. Mature ECP is cytotoxic to human bronchial epithelial (BEAS-2B) cells by specific binding to cell surface heparan sulfate proteoglycans (HSPGs) followed by endocytosis. ECP triggers apoptosis by caspase-8 activation through mitochondria-independent pathway. Increases in chromatin condensation, sub-G1 population, PARP cleavage, and DNA fragmentation indicate that ECP induces apoptosis in human bronchial epithelial (BEAS-2B) cells. |
| Immunogen | Recombinant protein within human Ribonuclease 3 aa 28-120. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG1 |
| Host Species | Mouse |
| Tested Applications | WBICCIHCFC |
| WB:1:500-1:2000: ICC:1:100: IHC:1:50-1:200: FC:1:50-1:100: |
|
| Species Reactivity | Human |
| Concentration | 2mg/ml |
| 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 |
|---|---|
| Molecular Weight(MW) | 18 kDa |
| Cellular Localization | Secreted. |
| SwissProt ID | P12724 |
|---|
Application
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.
Application
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).
Application
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).
Application
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.
Application
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.
Application
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.| Positive Control | U937, Hela, SHG-44, human spleen tissue, human kidney tissue. |
|---|---|
| Application Notes | WB:1:500-1:2000: ICC:1:100: IHC:1:50-1:200: FC:1:50-1:100: |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 50%Glycerol. Preservative: 0.05% Sodium Azide. |
