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Hersteller |
ProSci
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Kategorie |
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|
Typ |
Antibody Polyclonal |
|
Specific against |
Human, Mouse, Rat |
|
Format |
Liquid |
|
Applikationen |
WB, IF, IP, ELISA, IHC-P |
|
Menge |
0.02 mg |
|
Host |
Rabbit |
|
ArtNr |
PRS-2283-0.02mg |
|
eClass 6.1 |
32160702 |
|
eClass 9.0 |
32160702 |
|
Lieferbar |
 |
| Shipping |
blue ice or RT |
| By Research Area |
Apoptosis |
| Immunogen |
Anti-RIP3 antibody (2283) was raised against a peptide corresponding to 14 amino acids near the carboxy terminus of murine RIP3.
The immunogen is located within the last 50 amino acids of RIP3. |
| Applications |
WB: 0.1-0.5 μ g/mL; IHC-P: 5 μ g/mL; IF: 20 μ g/mL, IP: 20 μ g/mL.
Antibody validated: Western Blot in mouse and human samples; Immunohistochemistry in mouse and rat samples; Immunofluorescence in mouse and rat samples; Immunoprecipitation in human samples; Immunogold EM in mouse samples. All other applications and species not yet tested. |
| Positive Control 1 |
Cat. No. 1202 – A431 Cell Lysate |
| Positive Control 2 |
Cat. No. 1282 - 3T3/NIH Cell Lysate |
| Positive Control 3 |
Cat. No. 1285- C2C12 Cell Lysate |
| Positive Control 4 |
Cat. No. 1212- 3T3/Balb Cell Lysate |
| Predicted Molecular Weight |
Predicted: 53kD for mouse RIP3 and 57kD for human RIP3
Observed: 53kD for mouse RIP3 and 57kD for human RIP3 |
| Validation |
KO Validation (Figure 7, 9, 13) shows RIP3 expression detected by anti-RIP3 antibodies (2283) was disrupted in multiple tissues of RIP3 KO mice (figure 7, 13) and in RIP3 KO MEF cells (figure 9). KD validation (Figure 10): Anti-RIP3 antibody (2283) specificity was further verified by RIP3 specific knockdown. RIP3 signal in L929 cells transfected with RIP3 shRNA was disrupted in comparison with that in cells transfected with control shRNA. Regulated expression validation (Figure 7, 11, 13): RIP3 expression detected by anit-RIP3 antibodies (2283) was up-regulated by treatment of cerulein (figure 7), LPS (figure 11) or acetaminophen (figure 13) in different tissues or cells. Overexpression validation (Figure 12): RIP3 overexpression detected by anit-RIP3 antibodies (2283) was observed in three cell lines stably transfected with RIP3. |
| Isoforms |
Mouse RIP3 has one isoform(486aa, 53kD). Human RIP3 has 3 isoforms, including isoform 1 (518aa, 57kD), isoform 2 (252aa, 28kD) and isoform 3 (231aa, 25kD). Rat RIP3 also has one isoform (478aa, 52kD). 2283 can detect can detect isoforms of mouse and rat as well as human isoform 1. |
| Purification |
RIP3 Antibody is affinity chromatography purified via peptide column. |
| Clonality |
Polyclonal |
| Isotype |
IgG |
| Conjugate |
Unconjugated |
| Buffer |
RIP3 Antibody is supplied in PBS containing 0.02% sodium azide. |
| Concentration |
1 mg/ml |
| Storage Conditions |
RIP3 antibody can be stored at 4˚ C for three months and -20˚ C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures. |
| Related Products |
Cat. No. 2283P – RIP3 Peptide |
| Disclaimer |
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only. |
| Modifications |
None |
| Ncbi Official Symbol |
Ripk3 |
| Accession # |
AAF03133 |
| Protein Gi # |
6063101 |
| Ncbi Gene Id # |
56532 |
| User Note |
Optimal dilutions for each application to be determined by the researcher. |
| Ncbi Official Symbol |
Ripk3 |
| Ncbi Official Full Name |
receptor-interacting serine-threonine kinase 3 |
| Ncbi Organism |
Mus musculus |
| Swissprot # |
Q9QZL0 |
| Background |
RIP3 Antibody: Certain serine/threonine protein kinases, such as ASK1, RIP, DAP, and ZIP kinases, are mediators of apoptosis. Receptor interacting proteins including RIP and RIP2/RICK mediate apoptosis induced by TNFR1 and Fas, two prototype members in the death receptor family. A novel member in the RIP kinase family was recently identified and designated RIP3. RIP3 contains N-terminal kinase domain but, unlike RIP or RIP2, lacks the C-terminal death or CARD domain. RIP3 binds to RIP and TNFR1, mediates TNFR1 induced apoptosis, and attenuates RIP and TNFR1 induced NF-κ B activation. Overexpression of RIP3 induces apoptosis and NF-κ B activation. The messenger RNA of RIP3 is expressed in a subset of adult tissues. |
| Background References 1 |
Yu et al. Curr Biol. 1999; 9(10):539-42. |
| Background References 2 |
Sun et al. J Biol Chem. 1999; 274(24):16871-5. |
| Background References 3 |
Pazdernik et al. Mol Cell Bio. 1999; 19(10):6500-8 (WD0102) |
| Citation 1 |
He et al. Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-alpha. Cell. 2009; 137(6):1100-11. PMID: 19524512 |
| Citation 2 |
Zhang et al. Functional complementation between FADD and RIP1 in embryos and lymphocytes. Nature. 2011; 471(7338):373-6. PMID: 21368761 |
| Citation 3 |
Narayan et al. The NAD-dependent deacetylase SIRT2 is required for programmed necrosis. Nature. 2012; 492(7428):199-204. PMID: 23201684 |
| Citation 4 |
Li et al. Tissue damage negatively regulates LPS-induced macrophage necroptosis. Cell Death Differ. 2016; 23(9):1428-47. doi:10.1038/cdd.2016.21. Epub 2016. PMID: 26943325 |
| Citation 5 |
Yang et al. Regulation of RIP3 by the transcription factor Sp1 and the epigenetic regulator UHRF1 modulates cancer cell necroptosis. Cell Death Dis. 2017; 8(10):e3084. doi:10.1038/cddis.2017.483. PMID: 28981102 |
| Citation 6 |
Ramachandran et al. Receptor interacting protein kinase 3 is a critical early mediator of acetaminophen-induced hepatocyte necrosis in mice. Hepatology. 2013; 58(6):2099-108. PMID: 23744808 |
| Citation 7 |
Li et al. The RIP1/RIP3 necrosome forms a functional amyloid signaling complex required for programmed necrosis. Cell. 2012 ; 150(2):339-50. PMID: 22817896 |
| Citation 8 |
Vitner et al. RIPK3 as a potential therapeutic target for Gaucher's disease. Nat Med. 2014; 20(2):204-8. PMID: 24441827 |
| Citation 9 |
Wang et al. RNA viruses promote activation of the NLRP3 inflammasome through a RIP1-RIP3-DRP1 signaling pathway. Nat Immunol. 2014; 15(12):1126-33. PMID: 25326752 |
| Citation 10 |
Onizawa et al. The ubiquitin-modifying enzyme A20 restricts ubiquitination of the kinase RIPK3 and protects cells from necroptosis. Nat Immunol. 2015; 16(6):618-27. PMID: 25939025 |
| Citation 11 |
Murphy et al. The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism. Immunity. 2013; 39(3):443-53. PMID: 24012422 |
| Citation 12 |
Nogusa et al. RIPK3 Activates Parallel Pathways of MLKL-Driven Necroptosis and FADD-Mediated Apoptosis to Protect against Influenza A Virus. Cell Host Microbe 2016; 20(1):13-24. PMID: 27321907 |
| Citation 13 |
Pearson et al. EspL is a bacterial cysteine protease effector that cleaves RHIM proteins to block necroptosis and inflammation. Nat Microbiol. 2017 ; 2:16258. PMID: 28085133 |
| Citation 14 |
Yang et al. The end of RIPK1-RIPK3-MLKL-mediated necroptosis in acetaminophen-induced hepatotoxicity? Hepatology. 2016; 64(1):311-2PMID: 26418225 |
| Citation 15 |
Qu et al. Graphene oxide induces toll-like receptor 4 (TLR4)-dependent necrosisin macrophages. ACS Nano. 2013; 7(7):5732-45.PMID: 23734789 |
| Citation 16 |
Gandhirajan et al. Blockade of NOX2 and STIM1 signaling limits Lipo polysaccharide-induced vascular inflammation. J Clin Invest. 2013; 123(2):887-902. PMID: 23348743 |
| Citation 17 |
Fortes et al. Heme induces programmed necrosis on macrophages through autocrine TNF and ROS production. Blood. 2012; 119(10):2368-75. PMID: 22262768 |
| Citation 18 |
Xu et al. The cytoplasmic nuclear receptor RARγ controls RIP1 initiated cell death when cIAP activity is inhibited. Nat Commun. 2017; 8(1):425. PMID: 28871172 |
| Citation 19 |
Karunakaran et al. Targeting macrophage necroptosis for therapeutic and diagnostic interventions in atherosclerosis. Sci Adv. 2016 ; 2(7):e1600224. PMID: 27532042 |
| Citation 20 |
Chen et al. Mechanisms of necroptosis in T cells. J Exp Med. 2011 Apr 11; 208(4):633-41.PMID: 21402742 |
| 1st Image Caption |
Figure 1 Western Blot Validation in Human Cell Lines
Loading: 15 μ g of lysates per lane.Antibodies: RIP3 2283, (0.5 μ g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
| 2nd Image Caption |
Figure 2 Western Blot Validation in C2C12 Cells Mouse
Loading: 15 μ g of lysates per lane.Antibodies: RIP3 2283, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane 1: 2283, 0.1 μ g/mL in the presence of peptide blockingLane 2: 2283, 0.1 μ g/mLLane 3: 2283, 0.2 μ g/mLLane 4: 2283, 0.5 μ g/mL |
| 3rd Image Caption |
Figure 3 Western Blot Validation in Mouse Cell lines
Loading: 15 μ g of lysates per lane.Antibodies: RIP3 2283, (0.5 μ g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
| 4th Image Caption |
Figure 4 Immunohistochemistry Validation of RIP3 in Mouse Kidney Tissue
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-RIP3 antibody (2283) at 2.5 μ g/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚ C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
| 5th Image Caption |
Figure 5 Immunohistochemistry Validation of RIP3 in Rat Kidney Tissue
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-RIP3 antibody (2283) at 5 μ g/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚ C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
| 6th Image Caption |
Figure 6 Immunofluorescence Validation of RIP3 in Rat Kidney Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Kidney tissue labeling RIP3 with 2283 at 20 μ g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). |
| 7th Image Caption |
Figure 7 KO Validation of RIP3 in RIP3 KO Mice (He et al., 2009)
Western blot analysis of RIP3 with anti-RIP3 antibodies shows disrupted RIP3 expression in pancreas, liver spleen thymus and lung of RIP3 KO mice. Cerulein treatment upregulated RIP3 expression in the pancreas of WT mice. |
| 8th Image Caption |
Figure 8 Immunohistochemistry Validation of RIP3 in Fadd KO mice (Zhang et al., 2011)
RIP3 expression detected by anti-RIP3 antibodies (2283) was decreased in Fadd KO mice as compared to WT mice. |
| 9th Image Caption |
Figure 9 KO Validation of RIP3 in MEF Cells (He et al., 2012)
Western blot analysis of RIP3 with anti-RIP3 antibodies shows disrupted RIP3 expression in RIP3 KO MEFs, but not in WT MEF cells. |
| 10th Image Caption |
Figure 10 KD Validation of RIP3 in L929 Cells (Narayan et al., 2012)
Immunoblot analysis of RIP3 with anti-RIP3 antibodies (2283) shows RIP3 expression was disrupted in RIP3 knockdown L929 cells. |
| 11th Image Caption |
Figure 11 Regulated Expression Validation of RIP3 in Macrophages (Li et al., 2016)
Confocal microscopy shows colocalization of RIPK1 (green) and RIPK3 (red) in PMϕ and RIPK3 was detected by anti-RIPK3 antibodies (2283). PF or BCM suppressed LPS-induced upregulation of RIP3. |
| 12th Image Caption |
Figure 12 Overexpression Validation of RIP3 in Cancer Cell Lines (Yang et al., 2017)
The cancer cell lines were stably expressing flag-tagged RIP3 and RIP expression was detected by anti-RIP3 antibodies (2283) in RIP3-overexpressed cells. |
| 13th Image Caption |
Figure 13 Induced Expression of RIP3 by Acetaminophen in WT and RIP3 KO Mice (Ramachandran et al., 2013)
Wild-type and RIP3 KO mice were treated with300 mg/kg acetaminophen or saline. RIP3 expression detected by anti-RIP3 antibodies (2283) were up-regulated after acetaminophen treatment in the liver, while this effect was not observed in RIP3 KO mice. |
| 14th Image Caption |
Figure 14 IP Validation of RIP3 in HT29 Cells (Li et al., 2012)
RIP3 complexes isolated by immunoprecipitation with anti-RIP3 antibodies (2283) from HT-29 cells treated with T+Z+L after lysis in regular lysis buffer or buffer containing the indicated amount of urea or NaOH. |
| 15th Image Caption |
Figure 15 Immunogold EM Validation in MEF Cells (Li et al., 2012)
Clustering of RIP3 in necrotic MEFs shown by immunogold EM with anti-RIP3 antibodies (2283). Scale bars, 100 nm (RIP3). |
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Alle Produkte sind nur für Forschungszwecke bestimmt. Nicht für den menschlichen, tierärztlichen oder therapeutischen Gebrauch.
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