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RIP3 Polyclonal antibody

RIP3 Polyclonal Antibody for IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (3)

Applications

WB, IP, IHC, IF, CoIP, ELISA

Conjugate

Unconjugated

Cat no : 17563-1-AP

Synonyms

Receptor interacting protein 3, RIP 3, RIP like protein kinase 3, RIP3, RIPK3



Tested Applications

Positive WB detected inHT-29 cells, HeLa cells, SW 1990 cells, Jurkat cells, THP-1 cells, SGC-7901 cells
Positive IP detected inSW 1990 cells
Positive IHC detected inhuman pancreas tissue
Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:1000-1:4000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for IP and 1:200-1:1000 for WB
Immunohistochemistry (IHC)IHC : 1:100-1:400
Sample-dependent, check data in validation data gallery

Product Information

17563-1-AP targets RIP3 in WB, IP, IHC, IF, CoIP, ELISA applications and shows reactivity with human samples.

Tested Reactivity human
Cited Reactivityhuman, mouse, yeast, pig
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen RIP3 fusion protein Ag11759
Full Name receptor-interacting serine-threonine kinase 3
Calculated molecular weight 518 aa, 57 kDa
Observed molecular weight 57 kDa
GenBank accession numberBC062584
Gene symbol RIPK3
Gene ID (NCBI) 11035
Conjugate Unconjugated
Form Liquid
Purification Method Antigen affinity purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC 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.


Protocols

Product Specific Protocols
WB protocol for RIP3 antibody 17563-1-APDownload protocol
IHC protocol for RIP3 antibody 17563-1-APDownload protocol
IP protocol for RIP3 antibody 17563-1-APDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
humanWB

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
mouseIF

ACS Appl Mater Interfaces

Enhancing Tumor Therapy of Fe(III)-Shikonin Supramolecular Nanomedicine via Triple Ferroptosis Amplification

Authors - Wenjie Feng
mouseWB

Front Immunol

N-acetyl-L-cysteine alleviated the oxidative stress-induced inflammation and necroptosis caused by excessive NiCl2 in primary spleen lymphocytes

Authors - Xintong Zhang
humanWB

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
humanWB

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
mouse

Cell Mol Life Sci

Thioredoxin reductase 3 suppression promotes colitis and carcinogenesis via activating pyroptosis and necrosis.

Authors - Qi Liu