RIP3 Polyclonal antibody

RIP3 Polyclonal Antibody for WB, IP, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (3)

Applications

WB, IP, IF, IHC, 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, Jurkat cells, SW 1990 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 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)IHC : 1:100-1:400
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.

Product Information

17563-1-AP targets RIP3 in WB, IP, IF, IHC, 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
RRIDAB_2178659
Conjugate Unconjugated
Form Liquid
Purification MethodAntigen 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
humanWB

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
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