Phospho-MLKL (S345) Recombinant Rabbit Monoclonal Antibody [JM92-37]
cat.: ET1705-51
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse |
| Applications: | WB, IHC-P, IHC-Fr |
| Clonality: | Monoclonal |
| Clone number: | JM92-37 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 54 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser345 of mouse MLKL aa 314-363 / 472. |
| Positive control: | L929 treated with 20μM Z-VAD for 3.5 hours then added 100nM SM-164 and 20ng/ml TNF-α for 3 hours cell lysate, mouse spleen tissue, mouse lung tissue, mouse liver tissue, mouse colon tissue. |
| Subcellular location: | Cell membrane, Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IHC-P IHC-Fr |
1:1,000 1:50-1:100 1:100 |
| Uniprot #: | SwissProt: Q9D2Y4 Mouse |
| Alternative names: | 9130019I15Rik FLJ34389 hMLKL Mixed lineage kinase domain like Mixed lineage kinase domain like protein Mixed lineage kinase domain like pseudokinase Mixed lineage kinase domain-like protein Mlkl MLKL_HUMAN |
Images
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Fig1:
Western blot analysis of Phospho-MLKL (S345) on different lysates with Rabbit anti-Phospho-MLKL (S345) antibody (ET1705-51) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: L-929 cell lysate Lane 2: L929 treated with 20μM Z-VAD for 3.5 hours, then added 100nM SM-164 and 20ng/ml TNF-α for 3 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 54 kDa Exposure time: 1 minute 10 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 (ET1705-51) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-Phospho-MLKL (S345) antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-51, 1/50) 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. |
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Fig3: Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Phospho-MLKL (S345) antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-51, 1/50) 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. |
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Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Phospho-MLKL (S345) antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-51, 1/50) 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. |
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Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Phospho-MLKL (S345) antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-51, 1/50) 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. |
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Fig6:
Immunofluorescence analysis of frozen mouse spleen tissue with Rabbit anti-Phospho-MLKL (S345) antibody (ET1705-51) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1705-51, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.