RIPK3/RIP3 Antibody

Western Blot: RIPK3/RIP3 Antibody [NBP1-77299] - Analysis of RIP3 in HeLa cell lysate with RIP3 antibody at (A) 1 and (B) 2 ug/mL.

Immunohistochemistry: RIPK3/RIP3 Antibody [NBP1-77299] - IHC analysis of a murine kidney tissue section using RIP3 antibody at 20 ug/mL concentration and IF based detection.

Immunohistochemistry-Paraffin: RIPK3/RIP3 Antibody [NBP1-77299] - Rat kidney tissue with RIPK3 antibody at 5 ug/ml.

Immunocytochemistry/Immunofluorescence: RIPK3/RIP3 Antibody [NBP1-77299] - ICC/IF analysis of Rat's kidney cell culture using RIP3 antibody at 20 ug/mL concentration.

Immunohistochemistry-Paraffin: RIPK3/RIP3 Antibody [NBP1-77299] - Immunohistochemistry analysis of a paraffin embedded mouse kidney tissue section using RIP3 antibody at 2.5 ug/mL concentration.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ELISA, ICC/IF, IHC, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

RIPK3/RIP3 Antibody Summary

• Immunogen: Antibody was raised against a 14 amino acid peptide near the carboxy terminus of murine RIP3. The immunogen is located within the last 50 amino acids of RIP3.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: RIPK3
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/ml
  • ELISA 1:100 - 1:2000
  • Immunocytochemistry/Immunofluorescence 20 ug/ml
  • Immunohistochemistry 5 ug/ml
  • Immunohistochemistry-Paraffin 5 ug/ml
• Application Notes: This antibody can be used for detection of RIP3 by Western blot at 0.5 to 1 mg/ml. An approximately 57 kDa band can be detected.
• Theoretical MW: 57 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control Peptide:

  RIPK3/RIP3 Peptide (NBP1-77299PEP)

Reactivity Notes

Human reactivity reported in scientific literature (PMID: 26109138).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for RIPK3/RIP3 Antibody

• EC 2.7.11.1
• Receptor-interacting protein 3
• receptor-interacting serine/threonine-protein kinase 3
• receptor-interacting serine-threonine kinase 3
• RIP3
• RIP-3
• RIP3receptor interacting protein 3
• RIPK3
• RIP-like protein kinase 3

Background

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-kB activation. Overexpression of RIP3 induces apoptosis and NF-kappaB activation. The messenger RNA of RIP3 is expressed in a subset of adult tissues.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for RIPK3/RIP3 Antibody (NBP1-77299)(28)

Publications using NBP1-77299

• Rodriguez DA, Green DR. Generation and Use of Chimeric RIP Kinase Molecules to Study Necroptosis Methods Mol. Biol. Aug 24 2018 [PMID: 30136231] (WB, Mouse)
• Chen T, Pan H, Li J et al. Inhibiting of RIPK3 attenuates early brain injury following subarachnoid hemorrhage: Possibly through alleviating necroptosis Biomed. Pharmacother. Aug 13 2018 [PMID: 30114640] (ICC/IF, WB, Rat)
• Li P, Zhou Y, Goodwin AJ et al. Fli-1 Governs Pericyte Dysfunction in a Murine Model of Sepsis. J. Infect. Dis. Jul 20 2018 [PMID: 30053030] (WB, Mouse)
• Zamaraev AV, kopeina GS, Buchbinder JH et al. Caspase-2 is a negative regulator of necroptosis. Int. J. Biochem. Cell Biol. Jul 17 2018 [PMID: 30025878] (WB, Human)
• Piao X, Miura R, Miyake S et al. Blockade of TNF receptor superfamily 1 (TNFR1)-dependent and TNFR1-independent cell death is crucial for normal epidermal differentiation. J. Allergy Clin. Immunol. Mar 27 2018 [PMID: 29596938] (Mouse)
• Feldmann Friederike, Schenk Barbara, Martens Sofie et al. Sorafenib inhibits therapeutic induction of necroptosis in acute leukemia cells. Oncotarget 2017 [PMID: 28978109] (WB, Human)
• Martens S, Jeong M, Tonnus W et al. Sorafenib tosylate inhibits directly necrosome complex formation and protects in mouse models of inflammation and tissue injury Cell Death Dis 2017 Jun 29 [PMID: 28661484] (WB, Mouse)
• Sridharan H, Ragan KB, Guo H et al. Murine cytomegalovirus IE3-dependent transcription is required for DAI/ZBP1-mediated necroptosis EMBO Rep. 2017 Jun 12 [PMID: 28607035] (Mouse)
• Gong YN, Guy C, Olauson H et al. ESCRT-III Acts Downstream of MLKL to Regulate Necroptotic Cell Death and Its Consequences. Cell. Apr 6 2017 [PMID: 28388412] (Mouse)
• Ros U, Pena-Blanco A, Hanggi K et al. Necroptosis Execution Is Mediated by Plasma Membrane Nanopores Independent of Calcium. Cell Rep 2017 Apr 04 [PMID: 28380356] (WB, Mouse)
• Yang SH, Lee DK, Shin J et al. Nec-1 alleviates cognitive impairment with reduction of Abeta and tau abnormalities in APP/PS1 mice. EMBO Mol Med. Nov 17 2016 [PMID: 27861127] (IP, WB, Mouse)
• Shindo R, Yamazaki S, Ohmuraya M et al. Short form FLICE-inhibitory protein promotes TNFalpha-induced necroptosis in fibroblasts derived from CFLARs transgenic mice. Biochem. Biophys. Res. Commun. Nov 4 2016 [PMID: 27721066] (GS, Mouse)
• Qin D, Wang X, Li Y et al. MiR-223-5p and -3p Cooperatively Suppress Necroptosis in Ischemic/Reperfused Hearts. J. Biol. Chem. Aug 8 2016 [PMID: 27502281] (WB, Mouse)
• Seo J, Lee EW, Sung H et al. CHIP controls necroptosis through ubiquitylation- and lysosome-dependent degradation of RIPK3 Nat. Cell Biol. Mar 1 2016 [PMID: 26900751] (WB, IP, Human, Mouse)
• Wertz IE, Newton K, Seshasayee D et al. Phosphorylation and linear ubiquitin direct A20 inhibition of inflammation. Nature. 2015 Dec 17 [PMID: 26649818] (WB, Mouse)
• Yoon S, Bogdanov K, Kovalenko A, Wallach D et al. Necroptosis is preceded by nuclear translocation of the signaling proteins that induce it Cell Death Differ. 2015 Jul 17 [PMID: 26184911] (WB, Mouse)
• Miki Y, Akimoto J, Moritake K et al. Photodynamic therapy using talaporfin sodium induces concentration-dependent programmed necroptosis in human glioblastoma T98G cells Lasers Med Sci. 2015 Jun 25 [PMID: 26109138] (WB, Human)
  
  Details:
  human glioblastoma multiforma tumor cell line T98G treated with Control siRNA or (Fig. 4)
• Dara L, Johnson H, Suda J et al. Receptor Interacting protein kinase-1 mediates murine acetaminophen toxicity independent of the necrosome and not through necroptosis Hepatology. 2015 Jun 16 [PMID: 26077809] (WB, Mouse)
• Preyat N, Rossi M, Kers J et al. Intracellular nicotinamide adenine dinucleotide promotes TNF-induced necroptosis in a sirtuin-dependent manner. Cell Death Differ. Jan 1 2016 [PMID: 26001219] (WB, Human)
• Gautheron J, Vucur M, Reisinger F et al. A positive feedback loop between RIP3 and JNK controls non-alcoholic steatohepatitis. EMBO Mol Med. 2014 Jun 24 [PMID: 24963148] (WB, Mouse)
  
  Details:
  Figs 1B, S1, S11: RIPK3-/- and WT liver, Fig S10C: L929 fibrosarcoma cells. RIPK3 was detected between the 50 and 60 kDa marker. The specificity of the RIPK3 pAb was validated using RIPK3-/- and WT mouse liver by WB (Figs 1B, S11).
• Orozco S, Yatim N, Werner MR et al. RIPK1 both positively and negatively regulates RIPK3 oligomerization and necroptosis. Cell Death Differ. 2014 Jun 06 [PMID: 24902904] (WB, Mouse)
  
  Details:
  Figs 4b, S1B, S1I: Jaxson immortalized fibroblasts (Jax) and NIH3T3 cells transfected with RIPK3 fusion constructs (RIPK3-NIH3T3). RIPK3 was detected at ~50 kDa in Jax and at ~70 kDa in RIPK3-NIH3T3 cells. The specificity of the RIPK3 pAb was validated in RIPK3-NIH3T3 cells by WB (Figs 4b, S1B, S1I). Note: According to the authors, NIH3T3 cells do not expression endogenous RIPK3.
• Berger SB, Kasparcova V, Hoffman S et al. Cutting Edge: RIP1 kinase Activity Is Dispensable for Normal Development but Is a key Regulator of Inflammation in SHARPIN-Deficient Mice. J. Immunol. 2014 May 12 [PMID: 24821972] (WB, Mouse)
  
  Details:
  Bone marrow-derived macrophages, Fig 1C.
• Dillon CP, Weinlich R, Rodriguez DA et al. RIPk1 Blocks Early Postnatal Lethality Mediated by Caspase-8 and RIPk3. Cell. 2014 May 22 [PMID: 24813850] (WB, Mouse)
  
  Details:
  Mouse embryo fibroblasts, Figs 6D, 6H, S4D, S6A, S6D.
• Piao X, Komazawa-Sakon S, Nishina T et al. c-FLIP maintains tissue homeostasis by preventing apoptosis and programmed necrosis. Sci Signal. 2012 Dec 18 [PMID: 23250397] (WB)
  
  Details:
  WB: Fig 1B: primary RIPK3 knockout and wildtype mouse embryo fibroblasts (MEF) and 3T3-SA cells. Fig 2F: 3T3-SA Fig S1D: various tissues from RIPK3 knockout and wildtype mice, 3T3-SA Fig S3C: SVEC cells expressing RIPK3 siRNA or scrambled siRNA Fig SCD: N
• Dillon CP, Oberst A, Weinlich R et al. Survival function of the FADD-CASPASE-8-cFLIP(L) complex. Cell Rep. 2012 May 31 [PMID: 22675671]
• Oberst A, Dillon CP, Weinlich R et al. Catalytic activity of the caspase-8-FLIP(L) complex inhibits RIPK3-dependent necrosis. Nature. 2011 Mar 17 [PMID: 21368763]
• Kim JY, Kim YJ, Lee S, Park JH. BNip3 is a mediator of TNF-induced necrotic cell death. Apoptosis. 2011 Feb [PMID: 20963496] (WB)
  
  Details:
  RIP3 (IMG-5523-1 & 5523-2). WB: A549 cells treated or untreated with TNF-alpha, Fig 4D.
• Kaiser WJ, Upton JW, Mocarski ES. Receptor-interacting protein homotypic interaction motif-dependent control of NF-kappa B activation via the DNA-dependent activator of IFN regulatory factors. J Immunol. 2008 Nov 1 [PMID: 18941233] (WB, Human, Mouse)
  
  Details:
  WB [human 293T cells (2), mouse L929 cells (4)], Figs. 2A-C; 4B.

FAQs for RIPK3/RIP3 Antibody (NBP1-77299). (Showing 1 - 1 of 1 FAQs).

1. I am currently looking for a good antibody working as an ELISA capture antibody for mouse RIP3. Would you let me know if this application was tested during the development of your NovusBio (NBP1-77299) antibody.
   • Our product NBP1-77299 has been validated for use in ELISA. We tested this product in a direct ELISA format; it has not yet been tested in a sandwich ELISA format. The immunogen used to make this product falls between amino acids 473 to 486 of the mouse protein. If your detection antibody is outside of this range, it is possible that this combination will yield positive results.

Bioinformatics

• Gene Symbol: RIPK3
• Entrez:
  • 11035(Human)
  • 56532(Mouse)
  • 6063101(Mouse)
  • 246240(Rat)
• Uniprot:
  • AAF03133(Mouse)