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rabbit anti-RIP3 polyclonal antibody 3801

$445.00

Antibody summary

  • Rabbit polyclonal to RIP3
  • Suitable for: ELISA,IF,IHC-P,WB,IP
  • Isotype: IgG
  • 100 µg
SKU: 3801parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

RIP3

available sizes

100 µg

Available product – rabbit anti-RIP3 polyclonal antibody 3801

antibody
Tested applications
IHC,IHC,ICC/IF,ELISA
Recommended dilutions
Immunoblotting: use at 1ug/mL.

Positive control: NIH/3T3 cell lysate.

Immunohistochemistry: use at 5ug/mL.

These are recommended concentrations.

Enduser should determine optimal concentrations for their applications.
Immunogen
Synthetic peptide corresponding to aa 473-486 of mouse RIP3 (accession no. AAF03133).
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Storage buffer
PBS, pH 7.4.
Purity
peptide affinty purifcation
Clonality
polyclonal
Isotype
IgG
Compatible secondaries
goat anti-rabbit IgG, H&L chain specific, peroxidase conjugated, conjugated polyclonal antibody 9512
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody 2079
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody 7863
goat anti-rabbit IgG, H&L chain specific, Cross Absorbed polyclonal antibody 2371
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1715
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1720
Isotype control
Rabbit polyclonal - Isotype Control
target relevance
Protein names
Receptor-interacting serine/threonine-protein kinase 3 (EC 2.7.11.1) (RIP-like protein kinase 3) (Receptor-interacting protein 3) (RIP-3) (mRIP3)
Gene names
Ripk3,Ripk3 Rip3
Protein family
Protein kinase superfamily, TKL Ser/Thr protein kinase family
Mass
53322Da
Function
FUNCTION: Serine/threonine-protein kinase that activates necroptosis and apoptosis, two parallel forms of cell death (PubMed:27321907, PubMed:27746097, PubMed:27917412, PubMed:28607035, PubMed:32200799, PubMed:32296175). Necroptosis, a programmed cell death process in response to death-inducing TNF-alpha family members, is triggered by RIPK3 following activation by ZBP1 (PubMed:19590578, PubMed:22423968, PubMed:24012422, PubMed:24019532, PubMed:24557836, PubMed:27746097, PubMed:27819681, PubMed:27819682, PubMed:24095729, PubMed:32200799, PubMed:27321907, PubMed:32296175). Activated RIPK3 forms a necrosis-inducing complex and mediates phosphorylation of MLKL, promoting MLKL localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage (PubMed:24813849, PubMed:24813850, PubMed:27321907). In addition to TNF-induced necroptosis, necroptosis can also take place in the nucleus in response to orthomyxoviruses infection: following ZBP1 activation, which senses double-stranded Z-RNA structures, nuclear RIPK3 catalyzes phosphorylation and activation of MLKL, promoting disruption of the nuclear envelope and leakage of cellular DNA into the cytosol (PubMed:32200799, PubMed:32296175). Also regulates apoptosis: apoptosis depends on RIPK1, FADD and CASP8, and is independent of MLKL and RIPK3 kinase activity (PubMed:27321907). Phosphorylates RIPK1: RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation (By similarity). In some cell types, also able to restrict viral replication by promoting cell death-independent responses (PubMed:30635240). In response to flavivirus infection in neurons, promotes a cell death-independent pathway that restricts viral replication: together with ZBP1, promotes a death-independent transcriptional program that modifies the cellular metabolism via up-regulation expression of the enzyme ACOD1/IRG1 and production of the metabolite itaconate (PubMed:30635240). Itaconate inhibits the activity of succinate dehydrogenase, generating a metabolic state in neurons that suppresses replication of viral genomes (PubMed:30635240). RIPK3 binds to and enhances the activity of three metabolic enzymes: GLUL, GLUD1, and PYGL (By similarity). These metabolic enzymes may eventually stimulate the tricarboxylic acid cycle and oxidative phosphorylation, which could result in enhanced ROS production (By similarity). {ECO:0000250|UniProtKB:Q9Y572, ECO:0000269|PubMed:19590578, ECO:0000269|PubMed:22423968, ECO:0000269|PubMed:24012422, ECO:0000269|PubMed:24019532, ECO:0000269|PubMed:24095729, ECO:0000269|PubMed:24557836, ECO:0000269|PubMed:24813849, ECO:0000269|PubMed:24813850, ECO:0000269|PubMed:27321907, ECO:0000269|PubMed:27746097, ECO:0000269|PubMed:27819681, ECO:0000269|PubMed:27819682, ECO:0000269|PubMed:27917412, ECO:0000269|PubMed:28607035, ECO:0000269|PubMed:30635240, ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:32296175}.
Catalytic activity
CATALYTIC ACTIVITY: Reaction=ATP + L-seryl-[protein] = ADP + H(+) + O-phospho-L-seryl-[protein]; Xref=Rhea:RHEA:17989, Rhea:RHEA-COMP:9863, Rhea:RHEA-COMP:11604, ChEBI:CHEBI:15378, ChEBI:CHEBI:29999, ChEBI:CHEBI:30616, ChEBI:CHEBI:83421, ChEBI:CHEBI:456216; EC=2.7.11.1; Evidence={ECO:0000269|PubMed:10490590, ECO:0000269|PubMed:24095729, ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:32296175}; CATALYTIC ACTIVITY: Reaction=ATP + L-threonyl-[protein] = ADP + H(+) + O-phospho-L-threonyl-[protein]; Xref=Rhea:RHEA:46608, Rhea:RHEA-COMP:11060, Rhea:RHEA-COMP:11605, ChEBI:CHEBI:15378, ChEBI:CHEBI:30013, ChEBI:CHEBI:30616, ChEBI:CHEBI:61977, ChEBI:CHEBI:456216; EC=2.7.11.1; Evidence={ECO:0000269|PubMed:10490590, ECO:0000269|PubMed:24095729, ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:32296175};
Subellular location
SUBCELLULAR LOCATION: Cytoplasm, cytosol {ECO:0000269|PubMed:28607035, ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:32296175}. Nucleus {ECO:0000269|PubMed:28607035, ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:32296175}. Note=Mainly cytoplasmic (PubMed:32200799, PubMed:32296175). Present in the nucleus in response to influenza A virus (IAV) infection (PubMed:32200799). {ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:32296175}.
Tissues
TISSUE SPECIFICITY: Expressed in embryo and in adult spleen, liver, testis, heart, brain and lung. {ECO:0000269|PubMed:10490590}.
Structure
SUBUNIT: 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 (PubMed:27321907, PubMed:27819681, PubMed:28842570, PubMed:31519887). Interacts with MLKL; the interaction is direct and triggers necroptosis (PubMed:24012422, PubMed:27321907). Interacts with ZBP1 (via RIP homotypic interaction motif); interaction with ZBP1 activates RIPK3, triggering necroptosis (PubMed:19590578, PubMed:22423968, PubMed:27746097, PubMed:27819681, PubMed:27819682, PubMed:28607035, PubMed:32200799). 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 (By similarity). Binds TRAF2 and is recruited to the TNFR-1 signaling complex (By similarity). Interacts with PYGL, GLUL and GLUD1; these interactions result in activation of these metabolic enzymes (By similarity). Interacts with BIRC2/c-IAP1, BIRC3/c-IAP2 and XIAP/BIRC4 (By similarity). Interacts with ARHGEF2 (By similarity). Interacts with PELI1 (via atypical FHA domain); the phosphorylated form at Thr-187 binds preferentially to PELI1 (PubMed:29883609). Interacts with BUB1B, TRAF2 and STUB1 (By similarity). Interacts with CASP6 (By similarity). Component of the AIM2 PANoptosome complex, a multiprotein complex that drives inflammatory cell death (PANoptosis) (PubMed:34471287). {ECO:0000250|UniProtKB:Q9Y572, ECO:0000269|PubMed:19590578, ECO:0000269|PubMed:22423968, ECO:0000269|PubMed:24012422, ECO:0000269|PubMed:27321907, ECO:0000269|PubMed:27746097, ECO:0000269|PubMed:27819681, ECO:0000269|PubMed:27819682, ECO:0000269|PubMed:28607035, ECO:0000269|PubMed:28842570, ECO:0000269|PubMed:29883609, ECO:0000269|PubMed:31519887, ECO:0000269|PubMed:32200799, ECO:0000269|PubMed:34471287}.; SUBUNIT: (Microbial infection) Interacts (via RIP homotypic interaction motif) with murid herpesvirus protein RIR1; this interaction disrupts RIP3-RIP1 interactions characteristic of TNF-alpha induced necroptosis, thereby suppressing this death pathway. {ECO:0000269|PubMed:18442983, ECO:0000269|PubMed:30498077}.
Post-translational modification
PTM: RIPK1 and RIPK3 undergo reciprocal auto- and trans-phosphorylation (By similarity). Autophosphorylated following interaction with ZBP1 (PubMed:27819681). Phosphorylation of Ser-204 plays a role in the necroptotic function of RIPK3 (By similarity). Autophosphorylates at Thr-231 and Ser-232 following activation by ZBP1: phosphorylation at these sites is a hallmark of necroptosis and is required for binding MLKL (PubMed:23612963, PubMed:27819682). Phosphorylation at Thr-187 is important for its kinase activity, interaction with PELI1 and for its ability to mediate TNF-induced necroptosis (By similarity). {ECO:0000250|UniProtKB:Q9Y572, ECO:0000269|PubMed:23612963, ECO:0000269|PubMed:27819681, ECO:0000269|PubMed:27819682}.; PTM: 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 by STUB1 leading to its subsequent proteasome-dependent degradation. {ECO:0000250|UniProtKB:Q9Y572}.
Target Relevance information above includes information from UniProt accession : Q9QZL0
The UniProt Consortium

Data

benchmark-antibodies_anti-rip3_antibody_3801_1.gif
Western Blot Validation in Human Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: RIP3 3801, (0.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-rip3_antibody_3801_2.gif
Western Blot Validation in C2C12 Cells Mouse
Loading: 15 µg of lysates per lane. Antibodies: RIP3 3801, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane 1: 3801, 0.1 µg/mL in the presence of peptide blockingLane 2: 3801, 0.1 µg/mLLane 3: 3801, 0.2 µg/mLLane 4: 3801, 0.5 µg/mL
benchmark-antibodies_anti-rip3_antibody_3801_3.gif
Western Blot Validation in Mouse Cell lines
Loading: 15 µg of lysates per lane. Antibodies: RIP3 3801, (0.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-rip3_antibody_3801_4.gif
Immunohistochemistry Validation of RIP3 in Mouse Kidney Tissue
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-RIP3 antibody (3801) at 2.5 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-rip3_antibody_3801_5.jpg
Immunohistochemistry Validation of RIP3 in Rat Kidney Tissue
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-RIP3 antibody (3801) at 5 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-rip3_antibody_3801_6.jpg
Immunofluorescence Validation of RIP3 in Rat Kidney Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed Rat Kidney tissue labeling RIP3 with 3801 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
benchmark-antibodies_anti-rip3_antibody_3801_7.gif
KO Validation of RIP3 in RIP3 KO Mice (He et al., 2009)
Western blot analysis of RIP3 with anti-RIP3 antibodies shows disrupted RIP3 expression in pancreas, liver spleen thymus and lung of RIP3 KO mice. Cerulein treatment upregulated RIP3 expression in the pancreas of WT mice.
benchmark-antibodies_anti-rip3_antibody_3801_8.gif
Immunohistochemistry Validation of RIP3 in Fadd KO mice (Zhang et al., 2011)
RIP3 expression detected by anti-RIP3 antibodies (3801) was decreased in Fadd KO mice as compared to WT mice.
benchmark-antibodies_anti-rip3_antibody_3801_9.gif
KO Validation of RIP3 in MEF Cells (He et al., 2012)
Western blot analysis of RIP3 with anti-RIP3 antibodies shows disrupted RIP3 expression in RIP3 KO MEFs, but not in WT MEF cells.

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.




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Protocols

relevant to this product
Western blot
IHC
ICC

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