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RIP3 Antibody

RIP3 Polyclonal Antibody for IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IP, IHC, IF, 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 inJurkat cells, NIH/3T3 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 : 1:200-1:1000
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, ELISA applications and shows reactivity with human, mouse samples.

Reactivity human, mouse
Cited Species human, mouse
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen RIP3 fusion protein Ag11759
Full Name receptor-interacting serine-threonine kinase 3
Calculated molecular weight 518aa,57 kDa
Observed molecular weight 57 kDa
GenBank accession number BC062584
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.

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,IP

J Cell Physiol

MUC1 downregulation promotes TNF-α-induced necroptosis in human bronchial epithelial cells via regulation of the RIPK1/RIPK3 pathway.

Authors - Huojun Zhang
humanWB

Cardiovasc Drugs Ther

Dexmedetomidine Preconditioning Protects Cardiomyocytes Against Hypoxia/Reoxygenation-Induced Necroptosis by Inhibiting HMGB1-Mediated Inflammation.

Authors - Jingyi Chen
mouseWB,IHC

Cell Death Differ

The role of the LncRNA-FA2H-2-MLKL pathway in atherosclerosis by regulation of autophagy flux and inflammation through mTOR-dependent signaling.

Authors - Feng-Xia Guo
ratWB

Hum Gene Ther

A "Hibernating-like" viable state induced by Lentiviral Vector-Mediated PEDF overexpression in rat acute ischemic myocardium.

Authors - YanLiang Yuan
mouseWB

Toxins (Basel)

Evaluation of the Liver Toxicity of Pterocephalus hookeri Extract via Triggering Necrosis.

Authors - Rui Wang
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