Phospho-RIP3 (S232) Recombinant Rabbit Monoclonal Antibody [JE44-62]
cat.: HA721428
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse, Rat |
| Applications: | WB, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | JE44-62 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. 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: 53 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser232 of Human RIP3. |
| Positive control: | L-929 treated with 20μM Z-VAD for 3.5 hours add 100nM SM-164 and 20ng/ml TNF-α for 3 hours whole cell lysate, L-929 cells treated with 20μM Z-VAD for 2.75 hours, add 100nM SM-164 and 20ng/ml TNF-α for 2.25 hours. |
| Subcellular location: | Cytoplasm, cytosol, Nucleus. |
| Recommended Dilutions:
WB IF-Cell |
1:1,000 1:100 |
| Uniprot #: | SwissProt: Q9QZL0 Mouse | Q9Z2P5 Rat |
| Alternative names: | Receptor interacting protein 3 Receptor interacting serine threonine kinase 3 Receptor interacting serine/threonine protein kinase 3 Receptor-interacting protein 3 Receptor-interacting serine/threonine-protein kinase 3 RIP 3 RIP like protein kinase 3 RIP-3 RIP-like protein kinase 3 RIPK 3 RIPK3 RIPK3_HUMAN |
Images
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Fig1:
Western blot analysis of Phospho-RIP3 (S232) on different lysates with Rabbit anti-Phospho-RIP3 (S232) antibody (HA721428) at 1/1,000 dilution. Lane 1: L-929 whole cell lysate Lane 2: L-929 treated with 20μM Z-VAD for 3.5 hours, add 100nM SM-164 and 20ng/ml TNF-α for 3 hours whole cell lysate Lane 3: L-929 treated with λpp for 1 hour whole cell lysate Lane 4: L-929 treated with 20μM Z-VAD for 3.5 hours, add 100nM SM-164 and 20ng/ml TNF-α for 3 hours, treated with λpp for 1 hour whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 53 kDa Observed band size: 53 kDa Exposure time: 20 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 (HA721428) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of L-929 cells treated with or without 20μM Z-VAD for 2.75 hours, add 100nM SM-164 and 20ng/ml TNF-α for 2.25 hours, then treated with λpp for 1 hour labeling Phospho-RIP3 (S232) with Rabbit anti-Phospho-RIP3 (S232) antibody (HA721428) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-RIP3 (S232) antibody (HA721428) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.