RIPK3 Antibody - #DF10141

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製品: RIPK3 Antibody
カタログ: DF10141
タンパク質の説明: Rabbit polyclonal antibody to RIPK3
アプリケーション: WB IF/ICC
反応性: Human, Mouse
予測: Pig, Bovine, Horse, Sheep
分子量: 57 kDa; 57kD(Calculated).
ユニプロット: Q9Y572
RRID: AB_2840721

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MS Report
HPLC Report
MSDS
説明書
COA
プロトコル
WB Handbook
IHC Protocol
IF/ICC Protocol

製品説明

ソース:
Rabbit
アプリケーション:
WB 1:1000-3000, IF/ICC 1:100-1:500
*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.

反応性:
Human,Mouse
予測:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%)
クローナリティ:
Polyclonal
特異性:
RIPK3 Antibody detects endogenous levels of total RIPK3.
RRID:
AB_2840721
引用形式: Affinity Biosciences Cat# DF10141, RRID:AB_2840721.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
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.
別名:

折りたたみ／展開

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;

免疫原

免疫原:
Uniprot:

Q9Y572(RIPK3_HUMAN) >>Visit HPA database.

遺伝子(ID):
RIPK3(11035)
発現特異性:
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.

タンパク質配列:
MSCVKLWPSGAPAPLVSIEELENQELVGKGGFGTVFRAQHRKWGYDVAVKIVNSKAISREVKAMASLDNEFVLRLEGVIEKVNWDQDPKPALVTKFMENGSLSGLLQSQCPRPWPLLCRLLKEVVLGMFYLHDQNPVLLHRDLKPSNVLLDPELHVKLADFGLSTFQGGSQSGTGSGEPGGTLGYLAPELFVNVNRKASTASDVYSFGILMWAVLAGREVELPTEPSLVYEAVCNRQNRPSLAELPQAGPETPGLEGLKELMQLCWSSEPKDRPSFQECLPKTDEVFQMVENNMNAAVSTVKDFLSQLRSSNRRFSIPESGQGGTEMDGFRRTIENQHSRNDVMVSEWLNKLNLEEPPSSVPKKCPSLTKRSRAQEEQVPQAWTAGTSSDSMAQPPQTPETSTFRNQMPSPTSTGTPSPGPRGNQGAERQGMNWSCRTPEPNPVTGRPLVNIYNCSGVQVGDNNYLTMQQTTALPTWGLAPSGKGRGLQHPPPVGSQEGPKDPEAWSRPQGWYNHSGK

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
0
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9Y572 基板として

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 酵素として

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

研究背景

機能:

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.

細胞の位置付け:

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
組織特異性:

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.

サブユニット構造:

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).

タンパク質ファミリー:

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

研究領域

· 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)

参考文献

1). HNEAP Regulates Necroptosis of Cardiomyocytes by Suppressing the m5 C Methylation of Atf7 mRNA. Advanced science (Weinheim, Baden-Wurttemberg, Germany), 2023 (PubMed: 37870216) [IF=15.1]

Application: WB    Species: Mouse    Sample: cardiomyocytes

Figure 2 Knockdown of HNEAP attenuates H/R‐induced cardiomyocyte necroptosis. a–d) The isolated neonatal mice cardiomyocytes were transfected with HNEAP agomir (HNEAP) or its negative control (NC) for 24 h. a) The expression level of HNEAP was analyzed by qPCR (n = 6 independent experiments). b) Necroptosis was determined by the PI staining. DAPI indicates Nucleus. Bar = 25 µm. c) Quantitative analysis of the percentage of necroptotic cells (n = 6 independent experiments). d) The activity of lactate dehydrogenase (LDH) in cardiomyocytes after transfection with HNEAP or NC. (n = 6 independent experiments). e) The isolated neonatal mice cardiomyocytes were transfected with HNEAP antagomir (anta) or its negative control (anta‐NC) for 24 h. The expression level of HNEAP was analyzed by qPCR (n = 4 independent experiments). f–i) The isolated neonatal mice cardiomyocytes were transfected with HNEAP antagomir (anta) or its negative control (anta‐NC) for 24 h and then cells were treated with H/R as described in the Experimental Section. f) qPCR analysis of the expression level of HNEAP (n = 5 independent experiments). g) Necroptosis was determined by the PI staining. DAPI indicates Nucleus. Bar = 25 µm. h) Quantitative analysis of the percentage of necroptotic cells (n = 6 independent experiments). i) The upper panel shows the activity of LDH in cardiomyocytes after transfection with anta or anta‐NC and treated with H/R (n = 6 independent experiments). The lower panel is representative western blot showing the expression of RIPK1 and RIPK3. Data are presented as Mean ± SD. All data were analyzed using one‐way ANOVA.
2). Early Protection by Resveratrol in Rat Lung Transplantation. MEDICAL SCIENCE MONITOR, 2019 (PubMed: 30684444) [IF=3.1]

Application: IHC    Species: rat    Sample: lung

Figure 5.| Immunohistochemistry of RIPK3, MLKL, and TNF-a in lung grafts or in situ. The magnification of pictures is 400× (A). Protein expression was assessed as positive immune response or negative immune response depending on the immunohistochemistry score of different groups (NS group: n=8; DEM group: n=7; RES group: n=8). Positive immune response slices had more brown-staining cells in particular areas than the negative ones (B–D).

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