製品: RIPK1 Antibody
カタログ: AF7877
タンパク質の説明: Rabbit polyclonal antibody to RIPK1
アプリケーション: WB
反応性: Human, Mouse, Rat
予測: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
分子量: 76kDa; 76kD(Calculated).
ユニプロット: Q13546
RRID: AB_2844241

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製品説明

ソース:
Rabbit
アプリケーション:
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.

反応性:
Human,Mouse,Rat
予測:
Pig(86%), Bovine(86%), Horse(86%), Sheep(86%), Rabbit(86%), Dog(86%), Chicken(86%)
クローナリティ:
Polyclonal
特異性:
RIPK1 Antibody detects endogenous levels of total RIPK1.
RRID:
AB_2844241
引用形式: Affinity Biosciences Cat# AF7877, RRID:AB_2844241.
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

Cell death protein RIP; FLJ39204; OTTHUMP00000039163; Receptor (TNFRSF) interacting serine threonine kinase 1; receptor interacting protein 1; Receptor interacting protein; Receptor interacting protein kinase 1; Receptor interacting serine threonine protein kinase 1; Receptor TNFRSF interacting serine threonine kinase 1; Receptor-interacting protein 1; Receptor-interacting serine/threonine-protein kinase 1; Rinp; RIP 1; RIP; Rip-1; RIP1; RIPK 1; Ripk1; RIPK1_HUMAN; Serine threonine protein kinase RIP; Serine/threonine-protein kinase RIP;

免疫原

免疫原:
Uniprot:
遺伝子(ID):
タンパク質配列:
MQPDMSLNVIKMKSSDFLESAELDSGGFGKVSLCFHRTQGLMIMKTVYKGPNCIEHNEALLEEAKMMNRLRHSRVVKLLGVIIEEGKYSLVMEYMEKGNLMHVLKAEMSTPLSVKGRIILEIIEGMCYLHGKGVIHKDLKPENILVDNDFHIKIADLGLASFKMWSKLNNEEHNELREVDGTAKKNGGTLYYMAPEHLNDVNAKPTEKSDVYSFAVVLWAIFANKEPYENAICEQQLIMCIKSGNRPDVDDITEYCPREIISLMKLCWEANPEARPTFPGIEEKFRPFYLSQLEESVEEDVKSLKKEYSNENAVVKRMQSLQLDCVAVPSSRSNSATEQPGSLHSSQGLGMGPVEESWFAPSLEHPQEENEPSLQSKLQDEANYHLYGSRMDRQTKQQPRQNVAYNREEERRRRVSHDPFAQQRPYENFQNTEGKGTAYSSAASHGNAVHQPSGLTSQPQVLYQNNGLYSSHGFGTRPLDPGTAGPRVWYRPIPSHMPSLHNIPVPETNYLGNTPTMPFSSLPPTDESIKYTIYNSTGIQIGAYNYMEIGGTSSSLLDSTNTNFKEEPAAKYQAIFDNTTSLTDKHLDPIRENLGKHWKNCARKLGFTQSQIDEIDHDYERDGLKEKVYQMLQKWVMREGIKGATVGKLAQALHQCSRIDLLSSLIYVSQN

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
86
Horse
86
Bovine
86
Sheep
86
Dog
86
Chicken
86
Rabbit
86
Xenopus
67
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13546 基板として

Site PTM Type Enzyme
Phosphorylation
S6 Phosphorylation
K13 Ubiquitination
S14 Phosphorylation Q13546 (RIPK1)
S15 Phosphorylation Q13546 (RIPK1)
S20 Phosphorylation Q13546 (RIPK1)
S25 Phosphorylation
S32 Phosphorylation
T38 Phosphorylation
K49 Ubiquitination
K105 Ubiquitination
K115 Ubiquitination
K137 Ubiquitination
K140 Ubiquitination
K153 Ubiquitination
S161 Phosphorylation Q13546 (RIPK1)
K163 Ubiquitination
S166 Phosphorylation Q13546 (RIPK1)
K167 Ubiquitination
K184 Ubiquitination
K185 Ubiquitination
K284 Ubiquitination
S296 Phosphorylation
K302 Ubiquitination
S303 Phosphorylation
K306 Ubiquitination
S309 Phosphorylation
K316 Ubiquitination
S320 Phosphorylation
S330 Phosphorylation
S331 Phosphorylation
S333 Phosphorylation
T337 Phosphorylation
S346 Phosphorylation
K377 Ubiquitination
Y384 Phosphorylation
Y387 Phosphorylation
S389 Phosphorylation
S416 Phosphorylation
Y426 Phosphorylation
Y463 Phosphorylation
Y469 Phosphorylation
S470 Phosphorylation
S471 Phosphorylation
R477 Methylation
T483 Phosphorylation
R487 Methylation
Y490 Phosphorylation
K530 Acetylation
K571 Ubiquitination
K585 Ubiquitination
K596 Ubiquitination
K604 Ubiquitination
S610 Phosphorylation
K627 Ubiquitination
K642 Acetylation
K642 Ubiquitination
K648 Acetylation
S664 Phosphorylation

PTMs - Q13546 酵素として

Substrate Site Source
Q13233 (MAP3K1) S970 Uniprot
Q13546 (RIPK1) S14 Uniprot
Q13546 (RIPK1) S15 Uniprot
Q13546 (RIPK1) S20 Uniprot
Q13546 (RIPK1) S161 Uniprot
Q13546 (RIPK1) S166 Uniprot
Q5VWQ8 (DAB2IP) S728 Uniprot

研究背景

機能:

Serine-threonine kinase which is a key regulator of both cell death and cell survival. Exhibits kinase activity-dependent functions that trigger cell death and kinase-independent scaffold functions regulating inflammatory signaling and cell survival. Initiates ripoptocide which describes cell death that is dependent on RIPK1, be it apoptosis or necroptosis. Upon binding of TNF to TNFR1, RIPK1 is recruited to the TNF-R1 signaling complex (TNF-RSC also known as complex I) where it acts as a scaffold protein promoting cell survival, in part, by activating the canonical NF-kB pathway (By similarity). Specific conditions can however activate RIPK1, and its kinase activity then regulates assembly of two death-inducing complexes, namely complex IIa (RIPK1-FADD-CASP8) and the complex IIb (RIPK1-RIPK3-MLKL) and these complexes respectively drive apoptosis or necroptosis, a regulated form of necrosis. During embryonic development suppresses apoptosis and necroptosis and prevents the interaction of TRADD with FADD thereby limiting aberrant activation of CASP8 (By similarity). Phosphorylates DAB2IP at 'Ser-728' in a TNF- alpha-dependent manner, and thereby activates the MAP3K5-JNK apoptotic cascade. Required for ZBP1-induced NF-kappaB activation and activation of NF-kappaB by DNA damage and IR (By similarity).

PTMs:

Proteolytically cleaved by CASP8 at Asp-324. Cleavage is crucial for limiting apoptosis and necroptosis during embryonic development (By similarity). Cleavage abolishes NF-kappa-B activation and enhances the interaction of TRADD with FADD.

RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation. Phosphorylation of Ser-161 by RIPK3 is necessary for the formation of the necroptosis-inducing complex. Phosphorylation at Ser-25 represses its kinase activity and consequently prevents TNF-mediated RIPK1-dependent cell death. Phosphorylated at Ser-320 by MAP3K7 which requires prior ubiquitination with 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2 (By similarity). This phosphorylation positively regulates RIPK1 interaction with RIPK3 to promote necroptosis but negatively regulates RIPK1 kinase activity and its interaction with FADD to mediate apoptosis (By similarity).

Ubiquitinated with 'Lys-11'-, 'Lys-48'-, 'Lys-63'- and linear-linked type ubiquitin Ref.33). Polyubiquitination with 'Lys-63'-linked chains by TRAF2 induces association with the IKK complex. Deubiquitination of 'Lys-63'-linked chains and polyubiquitination with 'Lys-48'-linked chains by TNFAIP3 leads to RIPK1 proteasomal degradation and consequently down-regulates TNF-alpha-induced NFkappa-B signaling. 'Lys-48'-linked polyubiquitination by RFFL or RNF34 also promotes proteasomal degradation and negatively regulates TNF-alpha-induced NF-kappa-B signaling Ref.33). Linear polyubiquitinated; the head-to-tail linear polyubiquitination ('Met-1'-linked) is mediated by the LUBAC complex and decreases protein kinase activity. Deubiquitination of linear polyubiquitin by CYLD promotes the kinase activity (By similarity). Polyubiquitinated with 'Lys-48' and 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2, leading to activation of NF-kappa-B. Ubiquitinated with 'Lys-63'-linked chains by PELI1. Ubiquitination at Lys-377 with 'Lys-63'-linked chains by BIRC2/c-IAP1 and BIRC3/c-IAP2 is essential for its phosphorylation at Ser-320 mediated by MAP3K7 (By similarity). This ubiquitination is required for NF-kB activation, suppresses RIPK1 kinase activity and plays a critical role in preventing cell death during embryonic development (By similarity).

細胞の位置付け:

Cytoplasm. Cell membrane.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
サブユニット構造:

Homodimer. Interacts (via RIP homotypic interaction motif) with RIPK3 (via RIP homotypic interaction motif); this interaction induces RIPK1 phosphorylation and formation of a RIPK1-RIPK3 necroptosis-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 (via the death domain) with TNFRSF6 (via the death domain) and TRADD (via the death domain). Is recruited by TRADD to TNFRSF1A in a TNF-dependent process. Binds RNF216, EGFR, IKBKG, TRAF1, TRAF2 and TRAF3. Interacts with BNLF1. Interacts with SQSTM1 upon TNF-alpha stimulation. May interact with MAVS/IPS1. Interacts with ZFAND5. Interacts with RBCK1. Interacts with ZBP1 (By similarity). Interacts with BIRC2/c-IAP1, BIRC3/c-IAP2 and XIAP/BIRC4. Interacts (via kinase domain) with DAB2IP (via Ras-GAP domain); the interaction occurs in a TNF-alpha-dependent manner. Interacts with ARHGEF2. Interacts (via protein kinase domain) with RFFL; involved in RIPK1 ubiquitination. Interacts with RNF34; involved in RIPK1 ubiquitination (Ref.33). Interacts with TICAM1 and this interaction is enhanced in the presence of WDFY1. Interacts with PELI1. Interacts (via death domain) with CRADD (via death domain); the interaction is direct. Component of complex IIa composed of at least RIPK1, FADD and CASP8 (By similarity). Interacts with MAP3K7, CFLAR, CASP8, FADD and NEMO (By similarity).

(Microbial infection) Interacts with mumps virus protein SH; this interaction inhibits downstream NF-kappa-B pathway activation.

(Microbial infection) Interacts with Murid herpesvirus 1 protein RIR1.

タンパク質ファミリー:

The death domain mediates dimerization and activation of its kinase activity during necroptosis and apoptosis (PubMed:29440439). It engages other DD-containing proteins as well as a central (intermediate) region important for NF-kB activation and RHIM-dependent signaling (PubMed:10356400).

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

研究領域

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

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

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

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

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

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

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

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

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

参考文献

1). Extracellular vesicles of Fusobacterium nucleatum compromise intestinal barrier through targeting RIPK1-mediated cell death pathway. Gut Microbes, 2021 (PubMed: 33769187) [IF=12.2]

Application: WB    Species: human    Sample: Caco2 cells

Figure 2. |FnEVs promote TNF-α induced Caco2 cells death and barrier loss. (a) Diagram of groups. (b) Apoptosis was analyzed by using the annexin V FITC/PI assay. (c) Bar graphs show the relative mean of Caco2 cells apoptosis rate in different view fields. (d) The releasing levels of LDH were detected. (e) Representative images of TUNEL stainings of Caco-2 cells (green, TUNEL positive; blue, DAPI). Scale bar = 50 um. (f) Immunoblot analysis of protein extracts from Caco2 cells with the indicated antibodies

Application: IHC    Species: mouse    Sample: colon

Figure 6. |RIPK1 mediated epithelial cell death drives exacerbated barrier loss in FnEVs-treated colitis mice. (a) Representative images of TUNEL stainings of colon sections on day 3 after colitis induction (red, TUNEL positive; blue, DAPI). Scale bar = 50 um (up) or 20 um (down). (b) Representative images of immunohistochemical stainings of FADD, RIPK1 and cCASP3 in the colon sections on day 3 after colitis induction. Scale bar = 50 um or 20 um (downmost).

2). Sacubitril Ameliorates Cardiac Fibrosis Through Inhibiting TRPM7 Channel. Frontiers in Cell and Developmental Biology, 2021 (PubMed: 34778271) [IF=5.5]

3). Tanshinone I exerts cardiovascular protective effects in vivo and in vitro through inhibiting necroptosis via Akt/Nrf2 signaling pathway. Chinese Medicine, 2021 (PubMed: 34183021) [IF=4.9]

Application: WB    Species: Rat    Sample: H9c2 cells

Fig. 2 TI ameliorated t‑BHP induced cell necroptosis via RIP1/RIP3/MLKL pathway. a H9c2 cells were cultured with Nec-1 for 12 h. b Cells were exposed to t-BHP (150 μM) for 10 h after treated with Nec-1 for 2 h. c Cells were exposed to t-BHP (150 μM) for 10 h after treated with TI (1 μM) or Nec-1 (100 μM) for 2 h. MTT was employed to detect cell viability. d Cells were treated with t-BHP (150 μM) for 10 h when pretreated with TI (1 μM) or Nec-1 (100 μM) for 2 h, the LDH level was monitored by LDH kit. e–h Pretreated with TI (0.25, 0.5, and 1 μM) or Nec-1 (100 μM) for 2 h respectively, then exposed to t-BHP (150 μM) for 4 h, the protein expression was determined by western blotting. n = 3. *p < 0.05, **p < 0.01, ***p < 0.001 vs. t-BHP group

4). ERAD deficiency promotes mitochondrial dysfunction and transcriptional rewiring in human hepatic cells. JOURNAL OF BIOLOGICAL CHEMISTRY, 2020 (PubMed: 32978261) [IF=4.8]

5). Transient Inhibition of mTORC1 Signaling Ameliorates Irradiation-Induced Liver Damage. Frontiers in Physiology, 2019 (PubMed: 30984007) [IF=4.0]

Application: WB    Species: mouse    Sample: Liver

FIGURE 2 | Irradiation-induced cellular apoptosis in liver tissues. (A and B) Irradiated liver tissues were collected at days 1 and 3 after TBI. Representative pictures by TUNEL staining are shown (A, n = 6). Scale bar = 50 μm. Arrows indicate TUNEL-positive cells. The ratios of apoptotic cells in each field are presented in (B) (N.D: non-detected). (C and D) RIPK1 expression in liver tissues (n = 3). Expression of RIPK1 in liver tissues was detected by western blotting (C). GAPDH was used as a housekeeping control. Expression of RIPK1 was quantitated by ImageJ software (D).

6). Sugarcane leaf polysaccharide exerts a therapeutic effect on cardiovascular diseases through necroptosis. Heliyon, 2023 (PubMed: 38027563) [IF=4.0]

Application: WB    Species: Rat    Sample: H9c2 cells

Fig. 5 SLP alleviated TBHP-stimulated cell necroptosis through the RIP1/RIP3/MLKL signaling pathway in H9c2 cells. (A) H9c2 cells were incubated with SLP (25, 50, and 100 μg/mL) and then treated with TBHP (100 μM). The protein expression of p-RIP1, RIP1, p-RIP3, RIP3, p-MLKL, and MLKL was determined using western blotting. (B–D) Statistical analysis of the above protein expressions. n = 3. ##p < 0.01, ###p < 0.001, vs. the control group. *p < 0.05, **p < 0.01, ***p < 0.001, vs. TBHP group.

7). Metformin attenuates lung ischemia-reperfusion injury and necroptosis through AMPK pathway in type 2 diabetic recipient rats. BMC pulmonary medicine, 2024 (PubMed: 38745191) [IF=3.1]

Application: WB    Species: Rat    Sample:

Fig. 7 Metformin inhibited the necroptosis in type 2 diabetic lung transplant recipient rats. (A) RIPK1 representative western blot image. Full-length blots/gels are presented in Supplementary Fig. 2. (B) RIPK1 expression levels in lung grafts (n = 3). (C) RIPK3 representative western blot image. Full-length blots/gels are presented in Supplementary Fig. 3. (D) RIPK3 expression levels in lung grafts (n = 3). (E) MLKL representative western blot image. Full-length blots/gels are presented in Supplementary Fig. 4. (F) MLKL expression levels in lung grafts (n = 3). Compared with CS group, *P

8). Regulation of endoplasmic reticulum stress on autophagy and apoptosis of nucleus pulposus cells in intervertebral disc degeneration and its related mechanisms. PeerJ, 2024 (PubMed: 38666076) [IF=2.7]

Application: WB    Species: Human    Sample:

Figure 6 ERS affects the occurrence and development of IVDD by regulating programmed death. (A) ELISA detected the levels of IL-1β, TNF-α, iNOS and IL-6. (B) Western blot was used to detect the expressions of RIP1 and RIP3. The quantitative values are expressed as mean ± SD from at least three independent experiments

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