RIPK3 Polyclonal antibody

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  WB analysis using 17563-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 17563-1-AP (RIP3 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  WB analysis using 17563-1-AP

  SW 1990 cells were subjected to SDS PAGE followed by western blot with 17563-1-AP (RIP3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of Jurkat using 17563-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 17563-1-AP (RIP3 antibody at dilution of 1:300 incubated at room temperature for 1.5 hours.

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  IP experiment of SW 1990 using 17563-1-AP

  IP result of anti-RIP3 (IP:17563-1-AP, 3ug; Detection:17563-1-AP 1:300) with SW 1990 cells lysate 3000ug.

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  IHC staining of human placenta using 17563-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 17563-1-AP (RIP3 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human placenta using 17563-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 17563-1-AP (RIP3 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human pancreas using 17563-1-AP

  Immunohistochemical analysis of paraffin-embedded human pancreas tissue slide using 17563-1-AP (RIP3 antibody at dilution of 1:200 (under 10x lens).

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  IHC staining of human pancreas using 17563-1-AP

  Immunohistochemical analysis of paraffin-embedded human pancreas tissue slide using 17563-1-AP (RIP3 antibody at dilution of 1:200 (under 40x lens).

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  IF Staining of HT-29 using 17563-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HT-29 cells using RIP3 antibody (17563-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

RIPK3 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 17563-1-AP

Publications(68)

Host / Isotype

Rabbit / IgG

Reactivity

human and More (1)

Applications

WB, IHC, IF/ICC, IP, CoIP, ELISA

Synonyms

RIP3, EC:2.7.11.1, Receptor interacting protein 3, Receptor-interacting protein 3, Receptor-interacting serine/threonine-protein kinase 3

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Tested Applications

• Positive WB detected in: HT-29 cells, Jurkat cells, SW 1990 cells, THP-1 cells, SGC-7901 cells
• Positive IP detected in: SW 1990 cells
• Positive IHC detected in: human pancreas tissue, human placenta tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HT-29 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:100-1:400
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 60 publications below
• IHC: See 5 publications below
• IF: See 9 publications below
• IP: See 5 publications below
• CoIP: See 1 publications below

Product Information

17563-1-AP targets RIPK3 in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, yeast
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: RIPK3 fusion protein Ag11759
• Full Name: receptor-interacting serine-threonine kinase 3
• Calculated Molecular Weight: 518 aa, 57 kDa
• Observed Molecular Weight: 57-70 kDa
• GenBank Accession Number: BC062584
• Gene Symbol: RIPK3
• Gene ID (NCBI): 11035
• RRID: AB_2178659
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: Q9Y572
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Receptor-interacting protein 3 (RIP3, also known as RIPK3) is a serine-threonine protein involved in the regulation of inflammatory signaling and cell death. RIPK3, also named as RIP3, a Ser/Thr kinase of RIP (Receptor Interacting Protein) family, is a nucleocytoplasmic shuttling protein and its unconventional nuclear localization signal (NLS, 442-472 aa) is sufficient to trigger apoptosis in the nucleus (PMID: 18533105). It has 3 isoforms produced by alternative splicing.

What is the molecular weight of RIP3? Is RIP3 post-translationally modified?

The molecular weight of RIP3 is 57 kDa. During the induction of necroptosis, RIP3 migrates slower in SDS-PAGE due to its phosphorylation (PMID: 19524512). Additionally, RIP3 can be a subject of poly-ubiquitination, when targeted for degradation.

Are there any splice isoforms of RIP3?

Apart from full-length RIP3, there are two reported splice isoforms of RIP3: RIP3β and RIP3γ, and 28 and 25 kDa, respectively (PMID: 15896315).

What is the subcellular localization of RIP3?

RIP3 can shuttle between the cytoplasm and the nucleus. Although RIP3 forms necrosomes in the cytoplasm, a recent study suggests that the phosphorylation of RIP3, required for necroptosis, may also occur in the nucleus (PMID: 30271893). Additionally, the induction of necrosis by reactive oxygen species can cause transient translocation of RIP3 to mitochondria (PMID: 25206339).

What is the role of RIP3 in cell death (necroptosis)?

Activation of RIP3 kinase is required for the induction of necroptosis (PMID: 19524512, 19524513 and 19498109). Activation of RIP3 can be induced by interferons, death ligands, or by Toll-like receptors in response to pathogens. That leads to the phosphorylation of RIP3 and the formation of a β-amyloid-like protein complex. Phosphorylated RIP3 acts downstream by phosphorylation of MLKL (PMID: 30131615).

How to study necroptosis in a cell-based system?

The choice of the cell line is important. Many commonly used immortalized cell types are derived from cancers and may have very low RIL3 expression level (PMID: 25952668). Those cell lines are not going to be responsive to necrotic stimuli. A few of the examined cell types have high RIP3 levels: Jurkat, CCRF-CEM, U937, L929 cells, and mouse embryonic fibroblasts (PMID: 19524512). Good necroptosis readouts reflect an increased level of RIP3 protein and its phosphorylation.

Publications

Species	Application	Title
human	WB	Acta Pharm Sin B; Ligand-based substituent-anchoring design of selective receptor-interacting protein kinase 1 necroptosis inhibitors for ulcerative colitis therapy; Authors - Jing Zhu
human	WB	Cancer Lett; Lovastatin/SN38 co-loaded liposomes amplified ICB therapeutic effect via remodeling the immunologically-cold colon tumor and synergized stimulation of cGAS-STING pathway; Authors - Yi Yang
human	WB	Cancer Lett; NR4A1 depletion inhibits colorectal cancer progression by promoting necroptosis via the RIG-I-like receptor pathway; Authors - Jinghan Zhu
human	WB	Environ Pollut; Biotransformation of graphene oxide within lung fluids could intensify its synergistic biotoxicity effect with cadmium by inhibiting cellular efflux of cadmium.; Authors - Jianqiang Zhu
human	WB	Life Sci; TNF-α increases Staphylococcus aureus-induced death of human alveolar epithelial cell line A549 associated with RIP3-mediated necroptosis.; Authors - Shun-Hang Wen
human	WB	J Lipid Res; Hyperactivation of SREBP induces Pannexin-1-dependent lytic cell death; Authors - Yanni Xiong