Application
Fig1: Western blot analysis of PUMA on different lysates with Rabbit anti-PUMA antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HepG2 cell lysate, Lane 3: HDLM-2 cell lysate, Lane 4: NIH/3T3 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 24 seconds; 4-20% 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.Application
Fig2: ICC staining PUMA in Hela cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.Application
Fig3: ICC staining PUMA in SKOV-3 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.Application
Fig4: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue with Rabbit anti-PUMA antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour 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.Application
Fig5: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-PUMA antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour 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.Application
Fig6: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-PUMA antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour 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.Application
Fig7: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-PUMA antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.Application
Fig8: Flow cytometric analysis of Hela cells with PUMA antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.| Product Name | PUMA Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | The expression of PUMA is regulated by the tumor suppressor p53. PUMA is involved in p53-dependent and -independent apoptosis induced by a variety of signals, and is regulated by transcription factors, not by post-translational modifications. After activation, PUMA interacts with antiapoptotic Bcl-2 family members, thus freeing Bax and/or Bak which are then able to signal apoptosis to the mitochondria. Following mitochondrial dysfunction, the caspase cascade is activated ultimately leading to cell death. Several studies have shown that PUMA function is affected or absent in cancer cells. Additionally, many human tumors contain p53 mutations, which results in no induction of PUMA, even after DNA damage induced through irradiation or chemotherapy drugs.Other cancers, which exhibit overexpression of antiapotptic Bcl-2 family proteins, counteract and overpower PUMA-induced apoptosis. |
| Immunogen | Synthetic peptide within Human PUMA aa 153-193 / 193. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:500-1:2000 ICC/IF:1:50 IHC:1:1000-1:5000 FC:1:50 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | BBC 3 antibody Bbc3 antibody BBC3_HUMAN antibody BCL 2 binding component 3 antibody Bcl-2-binding component 3 antibody BCL2 binding component 3 antibody JFY 1 antibody JFY-1 antibody JFY1 antibody p53 up regulated modulator of apoptosis antibody p53 up-regulated modulator of apoptosis antibody p53 Upregulated Modulator of Apoptosis antibody PUMA alpha antibody PUMA/JFY1 antibody |
|---|---|
| Molecular Weight(MW) | 21kDa |
| Cellular Localization | Mitochondrion. |
Application
Fig1: Western blot analysis of PUMA on different lysates with Rabbit anti-PUMA antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HepG2 cell lysate, Lane 3: HDLM-2 cell lysate, Lane 4: NIH/3T3 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 24 seconds; 4-20% 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
Application
Fig2: ICC staining PUMA in Hela cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig3: ICC staining PUMA in SKOV-3 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Application
Fig4: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue with Rabbit anti-PUMA antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour 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.
Application
Fig5: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-PUMA antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour 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.
Application
Fig6: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-PUMA antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour 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.
Application
Fig7: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-PUMA antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.
Application
Fig8: Flow cytometric analysis of Hela cells with PUMA antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.| Application Notes | WB:1:500-1:2000 ICC/IF:1:50 IHC:1:1000-1:5000 FC:1:50 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
