CoraLite® Plus 488-conjugated RIP3 Polyclonal antibody

RIP3 Polyclonal Antibody for IF/ICC

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

IF/ICC

Conjugate

CoraLite® Plus 488 Fluorescent Dye

Cat no : CL488-17563

Synonyms

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

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Validation Data Gallery

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  IF Staining of HT-29 using CL488-17563

  Immunofluorescent analysis of (-20°C Methanol) fixed HT-29 cells using CoraLite® Plus 488 RIP3 antibody (CL488-17563) at dilution of 1:200.

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Tested Applications

• Positive IF/ICC detected in: HT-29 cells

Recommended dilution

• 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.

Product Information

CL488-17563 targets RIP3 in IF/ICC applications and shows reactivity with human samples.

• Tested Reactivity: human
• 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 Number: BC062584
• Gene Symbol: RIPK3
• Gene ID (NCBI): 11035
• Conjugate: CoraLite® Plus 488 Fluorescent Dye
• Excitation/Emission Maxima Wavelengths: 493 nm / 522 nm
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC 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.