Human Receptor-interacting serine/threonine-protein kinase 3(RIPK3) ELISA kit

Instructions
Code CSB-EL019737HU
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Receptor-interacting serine/threonine-protein kinase 3(RIPK3) ELISA Kit is designed to measure RIPK3 biological samples, including serum, plasma, tissue homogenates, and cell lysates, in a quantitative fashion. This kit has been verified by multiple tests with advantages of high sensitivity, strong specificity, precision less than 10%, and lot-to-lot consistency. Based on the Sandwich-ELISA technique in combination with the enzyme-substrate chromogenic reaction as well as colorimetric measurement, the levels of RIPK3 in the sample can be calculated.

RIPK3 is the essential regulator of necroptosis. The binding of TNF-α to TNF receptor 1 induces the activation of RIPK3, which recruits and phosphorylates downstream MLKL, finally leading to a cellular plasma membrane rupture and the leakage of cellular contents from dying cells that triggers an inflammatory response. The findings from Sarah Colijn etc. suggested that RIPK3 could play a novel, unexpected cell-type specific and athero-protective function.

Alternative Names Receptor interacting protein 3 ELISA Kit; Receptor interacting serine threonine kinase 3 ELISA Kit; Receptor interacting serine/threonine protein kinase 3 ELISA Kit; Receptor-interacting protein 3 ELISA Kit; Receptor-interacting serine/threonine-protein kinase 3 ELISA Kit; RIP 3 ELISA Kit; RIP like protein kinase 3 ELISA Kit; RIP-3 ELISA Kit; RIP-like protein kinase 3 ELISA Kit; RIPK 3 ELISA Kit; RIPK3 ELISA Kit; RIPK3_HUMAN ELISA Kit
Abbreviation RIPK3
Uniprot No. Q9Y572
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.156 ng/ml - 10 ng/ml
Sensitivity 0.039 ng/ml
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Materials provided
    • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human RIPK3 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
    • Two vials lyophilized standard --- Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
    • Biotin-labeled RIPK3 antibody (100 x concentrate) 1 x 120 μl ---Act as the detection antibody.
    • HRP-avidin (100 x concentrate) 1 x 120 μl --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
    • Biotin-antibody Diluent 1 x 15 ml ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
    • HRP-avidin Diluent 1 x 15 ml ---Dilute the high concentration HRP-avidin solution to an appropriate solution.
    • Sample Diluent 1 x 50 ml---Dilute the sample to an appropriate concentration.
    • Wash Buffer (25 x concentrate) 1 x 20 ml --- Wash away unbound or free substances.
    • TMB Substrate 1 x 10 ml --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
    • Stop Solution 1 x 10 ml --- Stop the color reaction. The solution color immediately turns from blue to yellow.
    • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
    • An instruction manual
Materials not provided
    • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
    • An incubator that can provide stable incubation conditions up to 37°C±5°C.
    • Centrifuge
    • Vortex
    • Squirt bottle, manifold dispenser, or automated microplate washer
    • Absorbent paper for blotting the microtiter plate
    • 50-300ul multi-channel micropipette
    • Pipette tips
    • Single-channel micropipette with different ranges
    • 100ml and 500ml graduated cylinders
    • Deionized or distilled water
    • Timer
    • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 7-14 working days

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 Q&A
Q:

Can I use this product CSB-EL019737HU to detece ripk3 in cell media?

A:
Thanks for your inquiry!
Sorry we usually test samples stated in the manual. We suggest you do a pretest if you want to test cell media

Target Background

Function
(From Uniprot)
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
Gene References into Functions
  1. The major function of RIP1 kinase activity in TNF-induced necroptosis is to autophosphorylate serine 161. This specific phosphorylation then enables RIP1 to recruit RIP3 and form a functional necrosome, a central controller of necroptosis. PMID: 28176780
  2. RIPK3-dependent cell death and inflammasome activation in FLT3-internal-tandem-duplication-expressing leukemia-initiating cells PMID: 27517160
  3. The necroptosis-inducing kinase RIPK3 reduces adipose tissue inflammation and glucose intolerance. PMID: 27323669
  4. We showed that RIP3 spontaneously drives a necroptosis-induced inflammation in established intestinal cell lines and in ileal/colonic samples from IBD patients. PMID: 28844856
  5. These data demonstrate that caspase-8 functions in synovial antigen-presenting cells to regulate the response to inflammatory stimuli by controlling RIPK3 action, and this delicate balance maintains homeostasis within the joint. PMID: 28978351
  6. The induced expression of RIP3 by UHRF1 RNAi depends on the presence of Sp1. Remarkably, the ectopic expression of RIP3 in RIP3-null cancer cells results in a decrease in tumor growth in mice. Therefore, our findings offer insights into RIP3 expression control in cancer cells and suggest an inhibitory effect of RIP3 on tumorigenesis. PMID: 28981102
  7. 2-hydroxyglutarate bound to DNMT1 and stimulated its association with the RIP3 promoter, inducing hypermethylation that reduces RIP3 protein and consequently impaired RIP3-dependent necroptosis. PMID: 28564603
  8. the in vivo effects were diametrically reversed with RIP3 deletion or RIP1 blockade, resulting in marked tumor protection. The dichotomy between the in vivo and in vitro results suggests that the microenvironmental milieu resulting from RIP1/RIP3 signaling is likely responsible for its protumorigenic effects PMID: 27932417
  9. Shikonin induces glioma cell necroptosis in vitro by reactive oxygen species overproduction and promoting RIP1/RIP3 necrosome formation. PMID: 28816233
  10. In critically ill trauma patients, plasma levels of the necroptosis mediator RIP3 at 48 h were associated with AKI stage and RBC transfusions. PMID: 26925809
  11. our results reveal that the necroptosis adaptor RIPK3 has key anti-inflammatory and anti-tumoral functions in the intestine, and define RIPK3 as a novel colon tumor suppressor PMID: 27344176
  12. adhesion-induced eosinophil cytolysis takes place through RIPK3-MLKL-dependent necroptosis, which can be counterregulated by autophagy PMID: 28412393
  13. Renal clear cell carcinoma cells cells express increased amounts of RIPK1 and RIPK3 and are poised to undergo necroptosis in response to TNFR1 signaling. PMID: 27362805
  14. The results highlight a new role of TSC2 in protecting glioblastoma against photodynamic therapy-induced cell death, and TSC2 and YWHAZ as new RIP3 partners. PMID: 27984090
  15. inactivation of RIP1/RIP3 resulted in reduction of SOCS1 protein levels and partial differentiation of AML cells. AML cells with inactivated RIP1/RIP3 signaling show increased sensitivity to IFN-gamma-induced differentiation. PMID: 27748372
  16. results reveal a pathway for MLKL-dependent programmed necrosis that is executed in the absence of RIPK3 and potentially drives the pathogenesis of severe liver diseases. PMID: 27756058
  17. Necroptosis signaling is modulated by the kinase RIPK1 and requires the kinase RIPK3 and the pseudokinase MLKL. (Review) PMID: 26865533
  18. Results demonstrate that RIPK3 restricts malignant myeloproliferation by activating the inflammasome, which promotes differentiation and cell death, and that loss of RIPK3 increases leukemic burden in mice. Reduced RIPK3 expression is observed across several human acute myeloid leukemia subtypes. PMID: 27411587
  19. Data identify RIPK3 and the inflammasome as key tumor suppressors in acute myeloid leukemia (AML). PMID: 27411587
  20. The main route of cell death induced by shikonin is RIP1K-RIP3K-mediated necroptosis. PMID: 26496737
  21. Results suggest that impaired hepatic proteasome function by alcohol exposure may contribute to hepatic accumulation of RIP3 resulting in necroptosis and steatosis while RIP1 kinase activity is important for alcohol-induced inflammation. PMID: 26769846
  22. The expression level of RIP3K was significantly lower in the malignant tumors. PMID: 26749282
  23. CNOT3 suppression promotes necroptosis by stabilizing mRNAs for cell death-inducing proteins, Ripk1 and Ripk3. PMID: 26437789
  24. RIPK3 expression may allow unmasking the necroptotic signalling machinery in melanoma and points to reactivation of this pathway as a treatment option for metastatic melanoma. PMID: 26355347
  25. Additionally, later in infection, RIP3 is cleaved by the coxsackievirus B3-encoded cysteine protease 3C(pro), which serves to abrogate RIP3-mediated necrotic signaling and induce a nonnecrotic form of cell death. PMID: 26269957
  26. although JNK activation and RIP3 expression are induced by FS, neither contributes to the liver injury. PMID: 25423287
  27. The RIP3 expression is reduced in tumors compared to normal tissue in 85% of breast cancer patients, suggesting that RIP3 deficiency is positively selected during tumor growth/development. PMID: 25952668
  28. Data implicate the infiltrating macrophages as a source of damaging inflammasomes after photoreceptor detachment in a RIP3-dependent manner and suggest a novel therapeutic target for treatment of retinal diseases. PMID: 25906154
  29. PolyIC stimulation of cervical cancer cells induced necroptotic cell death, which was strictly dependent on the expression of the receptor-interacting protein kinase RIPK3. PMID: 25888634
  30. Data suggest that neoalbaconol-induced necroptosis include receptor interacting serine/threonine kinase 1-dependent expression of tumor necrosis factor alpha and receptor interacting serine/threonine kinase 3-dependent generation of reactive oxygen species. PMID: 25575821
  31. Herpes simplex virus 1- and 2 ICP6 and ICP10 proteins prevent necroptosis in human cells by inhibiting the interaction between receptor-interacting protein kinase 1 (RIP1) and RIP3, a key step in tumor necrosis factor (TNF)-induced necroptosis. PMID: 25674983
  32. High expression of RIP3 in keratinocytes from toxic epidermal necrolysis patients potentiates MLKL phosphorylation/activation and necrotic cell death. PMID: 25748555
  33. RIP3 silencing in leukemia cells results in suppression of the complex regulation of the apoptosis/necroptosis switch and NF-kappaB activity. PMID: 25144719
  34. Suppression of RIP3-dependent necroptosis by human cytomegalovirus PMID: 25778401
  35. RIP3 activation following the induction of necroptosis requires the activity of an HSP90 and CDC37 cochaperone complex. PMID: 25852146
  36. The results of this study showed that cerebral ischemia activates transcriptional changes that lead to an increase in the endogenous RIP3 protein level. PMID: 24746856
  37. Enhanced RIP3 signaling in aneurysmal tissues contributes to abdominal aortic aneursym progression by causing smooth muscle cell necroptosis, as well as stimulating vascular inflammation. PMID: 25563840
  38. RIP3-dependent necroptosis mediates non-alcoholic steatohepatitis-induced liver fibrosis via activation of JNK, MCP-1-mediated recruitment of monocytes, and an expansion of intrahepatic biliary/progenitor cells. PMID: 24963148
  39. RIPK3 serves as a negative regulator of selective autophagy by regulating regulates p62-LC3 complex formation via the caspase-8-dependent cleavage of p62 PMID: 25450619
  40. RIP3 holds both necroptosis and apoptosis in balance through a Ripoptosome-like platform. PMID: 25459880
  41. an alanine residue substitution for Ser(89) enhanced RIP1 kinase activity and TNF-induced programmed necrosis without affecting RIP1-RIP3 necrosome formation. PMID: 24059293
  42. targeting the RIP kinase signalling pathway could be an effective therapeutic intervention in retinal degeneration patients. PMID: 24413151
  43. RIP3-mediated MLKL phosphorylation, though important for downstream signaling, is dispensable for stable complex formation between RIP3 and MLKL. PMID: 24095729
  44. Report role of MLKL/RIP3 pathway in necrotic membrane disruption. PMID: 24703947
  45. RIP3 protein expression is significantly increased in the inflamed tissue of inflammatory boowel disease pediatric patients. PMID: 24322838
  46. The protein levels of crucial modulators of necroptosis, RIP1 and RIP3, are increased by shikonin treatment in primary tumor tissues. PMID: 24314238
  47. the importance of the RIP3-MLKL interaction in the formation of functional necrosomes and suggest that translocation of necrosomes to mitochondria-associated membranes is essential for necroptosis signaling. PMID: 23612963
  48. procaspase-8 activity is essential for cell survival by inhibiting both apoptotic and nonapoptotic cell death dependent on receptor-interacting protein kinase 1 (RIP1) and RIP3 PMID: 23071110
  49. Study shows that RIP1 and RIP3 form an amyloid structure through their RIP homotypic interaction motifs and that this heterodimeric amyloid structure is a functional signaling complex that mediates programmed necrosis. PMID: 22817896
  50. study suggests that MLKL is a key RIP3 downstream component of TNF-induced necrotic cell death PMID: 22421439

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Subcellular Location Cytoplasm, cytosol, Cell membrane, Mitochondrion
Protein Families Protein kinase superfamily, TKL Ser/Thr protein kinase family
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.
Database Links

HGNC: 10021

OMIM: 605817

KEGG: hsa:11035

STRING: 9606.ENSP00000216274

UniGene: Hs.268551

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