RIPK3 Antibody - #AF4808

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Product: RIPK3 Antibody
Catalog: AF4808
Description: Rabbit polyclonal antibody to RIPK3
Application: WB
Reactivity: Human
Mol.Wt.: 46-62kDa; 57kD(Calculated).
Uniprot: Q9Y572
RRID: AB_2844793

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human
Clonality:
Polyclonal
Specificity:
RIPK3 Antibody detects endogenous levels of total RIPK3.
RRID:
AB_2844793
Cite Format: Affinity Biosciences Cat# AF4808, RRID:AB_2844793.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Receptor interacting protein 3; Receptor interacting serine threonine kinase 3; Receptor interacting serine/threonine protein kinase 3; Receptor-interacting protein 3; Receptor-interacting serine/threonine-protein kinase 3; RIP 3; RIP like protein kinase 3; RIP-3; RIP-like protein kinase 3; RIPK 3; RIPK3; RIPK3_HUMAN;

Immunogens

Immunogen:
Uniprot:

Q9Y572(RIPK3_HUMAN) >>Visit HPA database.

Gene(ID):
RIPK3(11035)
Expression:
Q9Y572 RIPK3_HUMAN:

Highly expressed in the pancreas. Detected at lower levels in heart, placenta, lung and kidney. Isoform 3 is significantly increased in colon and lung cancers.

Sequence:
MSCVKLWPSGAPAPLVSIEELENQELVGKGGFGTVFRAQHRKWGYDVAVKIVNSKAISREVKAMASLDNEFVLRLEGVIEKVNWDQDPKPALVTKFMENGSLSGLLQSQCPRPWPLLCRLLKEVVLGMFYLHDQNPVLLHRDLKPSNVLLDPELHVKLADFGLSTFQGGSQSGTGSGEPGGTLGYLAPELFVNVNRKASTASDVYSFGILMWAVLAGREVELPTEPSLVYEAVCNRQNRPSLAELPQAGPETPGLEGLKELMQLCWSSEPKDRPSFQECLPKTDEVFQMVENNMNAAVSTVKDFLSQLRSSNRRFSIPESGQGGTEMDGFRRTIENQHSRNDVMVSEWLNKLNLEEPPSSVPKKCPSLTKRSRAQEEQVPQAWTAGTSSDSMAQPPQTPETSTFRNQMPSPTSTGTPSPGPRGNQGAERQGMNWSCRTPEPNPVTGRPLVNIYNCSGVQVGDNNYLTMQQTTALPTWGLAPSGKGRGLQHPPPVGSQEGPKDPEAWSRPQGWYNHSGK

PTMs - Q9Y572 As Substrate

Site PTM Type Enzyme
K5 Ubiquitination
K55 Ubiquitination
T182 Phosphorylation
S199 Phosphorylation Q9Y572 (RIPK3)
T224 Phosphorylation
S227 Phosphorylation Q9Y572 (RIPK3)
S241 Phosphorylation
T252 Phosphorylation
S299 Phosphorylation
T300 Phosphorylation
S316 Phosphorylation
S320 Phosphorylation
T325 Phosphorylation
S339 Phosphorylation
S359 Phosphorylation
K363 Ubiquitination
S372 Phosphorylation
S410 Phosphorylation
T412 Phosphorylation
R486 Methylation

PTMs - Q9Y572 As Enzyme

Substrate Site Source
Q8NB16 (MLKL) T357 Uniprot
Q8NB16 (MLKL) S358 Uniprot
Q9UER7 (DAXX) S668 Uniprot
Q9Y572 (RIPK3) S199 Uniprot
Q9Y572 (RIPK3) S227 Uniprot

Research Backgrounds

Function:

Essential for necroptosis, a programmed cell death process in response to death-inducing TNF-alpha family members. Upon induction of necrosis, RIPK3 interacts with, and phosphorylates RIPK1 and MLKL to form a necrosis-inducing complex. RIPK3 binds to and enhances the activity of three metabolic enzymes: GLUL, GLUD1, and PYGL. These metabolic enzymes may eventually stimulate the tricarboxylic acid cycle and oxidative phosphorylation, which could result in enhanced ROS production.

PTMs:

RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation. Phosphorylation of Ser-199 plays a role in the necroptotic function of RIPK3. Phosphorylation at Ser-227 is required for binding MLKL. Phosphorylation at Thr-182 is important for its kinase activity, interaction with PELI1 and PELI1-mediated 'Lys-48'-linked polyubiquitination and for its ability to mediate TNF-induced necroptosis.

Polyubiquitinated with 'Lys-48' and 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2, leading to activation of NF-kappa-B. Polyubiquitinated with 'Lys-48'-linked chains by PELI1 leading to its subsequent proteasome-dependent degradation. Ubiquitinated by STUB1 leading to its subsequent proteasome-dependent degradation.

Subcellular Location:

Cytoplasm>Cytosol. Cell membrane. Mitochondrion.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Highly expressed in the pancreas. Detected at lower levels in heart, placenta, lung and kidney. Isoform 3 is significantly increased in colon and lung cancers.

Subunit Structure:

Interacts (via RIP homotypic interaction motif) with RIPK1 (via RIP homotypic interaction motif); this interaction induces RIPK1 phosphorylation and formation of a RIPK1-RIPK3 necrosis-inducing complex. Upon TNF-induced necrosis, the RIPK1-RIPK3 dimer further interacts with PGAM5 and MLKL; the formation of this complex leads to PGAM5 phosphorylation and increase in PGAM5 phosphatase activity. Interacts with MLKL; the interaction is direct. Binds TRAF2 and is recruited to the TNFR-1 signaling complex. Interacts with PYGL, GLUL and GLUD1; these interactions result in activation of these metabolic enzymes. Interacts with BIRC2/c-IAP1, BIRC3/c-IAP2 and XIAP/BIRC4. Interacts with ARHGEF2. Interacts with PELI1 (via atypical FHA domain); the phosphorylated form at Thr-182 binds preferentially to PELI1. Interacts with BUB1B, TRAF2 and STUB1. Interacts with ZBP1 (By similarity).

Family&Domains:

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.   (View pathway)

References

1). Analysis of multiple programmed cell death-related prognostic genes and functional validations of necroptosis-associated genes in oesophageal squamous cell carcinoma. EBioMedicine, 2024 (PubMed: 38101299) [IF=11.1]
2). Quercetin-induced pyroptosis in colon cancer through NEK7-mediated NLRP3 inflammasome-GSDMD signaling pathway activation. American journal of cancer research, 2024 (PubMed: 38590424) [IF=5.3]

Application: WB    Species: Human    Sample: HCT116 and HT29 cells

Figure 4 Que-induced cell death independent of necroptosis pathway activation. A. Protein levels of MLKL and p-MLKL in HCT116 and HT29 cells treated with various doses of GSK’872 were measured using western blotting assay (n=3, where “n” represents the number of independent repeated experiments). Values are presented as mean ± SD. *P

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