Name: Recombinant Mouse Mixed lineage kinase domain-like protein (Mlkl)
Brand: CUSABIO
SKU: CSB-YP861529MO

Product Details

Purity

Greater than 90% as determined by SDS-PAGE.

Target Names

Mlkl

Uniprot No.

Q9D2Y4

Research Area

Others

Alternative Names

MlklMixed lineage kinase domain-like protein

Species

Mus musculus (Mouse)

Source

Yeast

Expression Region

1-472aa

Target Protein Sequence

MDKLGQIIKLGQLIYEQCEKMKYCRKQCQRLGNRVHGLLQPLQRLQAQGKKNLPDDITAALGRFDEVLKEANQQIEKFSKKSHIWKFVSVGNDKILFHEVNEKLRDVWEELLLLLQVYHWNTVSDVSQPASWQQEDRQDAEEDGNENMKVILMQLQISVEEINKTLKQCSLKPTQEIPQDLQIKEIPKEHLGPPWTKLKTSKMSTIYRGEYHRSPVTIKVFNNPQAESVGIVRFTFNDEIKTMKKFDSPNILRIFGICIDQTVKPPEFSIVMEYCELGTLRELLDREKDLTMSVRSLLVLRAARGLYRLHHSETLHRNISSSSFLVAGGYQVKLAGFELSKTQNSISRTAKSTKAERSSSTIYVSPERLKNPFCLYDIKAEIYSFGIVLWEIATGKIPFEGCDSKKIRELVAEDKKQEPVGQDCPELLREIINECRAHEPSQRPSVDGRSLSGRERILERLSAVEESTDKKV
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.

Mol. Weight

56.3kDa

Protein Length

Full Length

Tag Info

N-terminal 6xHis-tagged

Form

Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Customer Reviews and Q&A

■ Customer Reviews

Average Rating:

5.0 - 1 reviews

Submit a Review here

Applications : In vitro kinase assays

Review: In vitro kinase reactions of purified OASL–GFP, GST–RIPK3, ZBP1–BFP and MLKL. OASL–RIPK3–ZBP1 were induced for phase separation, subjected to kinase reaction and immunoblotted for Ser227 autophosphorylation of RIPK3 and Ser358 phosphorylation of MLKL (asterisk indicates a nonspecific band).

By Anonymous

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■ Q&A

We would like your protein CSB-YP861529MO at a higher concentration than you usually supply. Is this something that Cusabio is able to do? If so, what concentrations are available and what is the pricing? Thank you very much for your time and assistance.

Thanks for your inquiry. The concentration of this protein delivered before is generally 0.1-0.5mg/ml, and once it is 1mg/ml. The conventional concentration of Yeast expressed proteins is 0.1-2mg/ml.
Please confirm your concentration demand first and we will confirm whether we could satisfy your demand. Please remark this request when sending your order.

Target Background

Function

Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process. Does not have protein kinase activity. Activated following phosphorylation by RIPK3, leading to homotrimerization, localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage. 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. Binds to highly phosphorylated inositol phosphates such as inositolhexakisphosphate (InsP6) which is essential for its necroptotic function.

Gene References into Functions

Aldehyde dehydrogenase 2 deficiency negates chronic low-to-moderate alcohol consumption-induced cardioprotecion possibly via ROS-dependent apoptosis and RIP1/RIP3/MLKL-mediated necroptosis. PMID: 27840306
M. tuberculosis and TNFalpha synergise to induce necroptosis in murine fibroblasts via RIPK1-dependent mechanisms and characterized by phosphorylation of Ser345 of the MLKL necroptosis death effector. PMID: 28892415
hydrostatic pressure (EHP)-induced RGC-5 cell necroptosis was mediated by MLKL PMID: 28981598
Biological events and molecular signaling following MLKL activation during necroptosis have been reported. PMID: 28854080
The authors report here that male reproductive organs of both Ripk3- and Mlkl-knockout mice retain 'youthful' morphology and function into advanced age, while those of age-matched wild-type mice deteriorate. Feeding of wild-type mice with an RIPK1 inhibitor prior to the normal onset of age-related changes in their reproductive organs blocked the appearance of signs of aging. PMID: 28807105
Data indicate that mixed-lineage kinase domain-like protein (MLKL) oligomerization, membrane translocation, and cell death occur simultaneously with NLR family pyrin domain containing 3 (NLRP3) activation in bone marrow-derived macrophages. PMID: 28096356
Pull down experiments with biotinylated Sorafenib show that it binds independently RIPK1, RIPK3 and MLKL. Moreover, it inhibits RIPK1 and RIPK3 kinase activity. In vivo Sorafenib protects against TNF-induced systemic inflammatory response syndrome (SIRS) and renal ischemia-reperfusion injury (IRI). PMID: 28661484
Knock-out mice deficient in MLKL or RIP3 had increased survival and reduced pulmonary injury during Serratia marcescens pneumonia. PMID: 28387756
Our findings reveal that MLKL and FADD play critical roles in preventing lymphoproliferative disease and activating the NLRP3 inflammasome PMID: 27498868
RIPK3 and/or MLKL may exert functions independently of necroptosis. PMID: 27523270
Data suggest that necroptotic cells externalize phosphatidylserine (PS) after translocation of phosphorylated Mlkl to cell membrane; necroptotic cells with exposed PS release extracellular vesicles containing Mlkl; inhibition of Mlkl after PS exposure can reverse process of necroptosis and restore cell viability. PMID: 28650960
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
Mice deficient in RIPK3 or doubly deficient in MLKL and FADD, but not MLKL alone, are more susceptible to influenza A virus than their wild-type counterparts, revealing an important role for RIPK3-mediated apoptosis in antiviral immunity. PMID: 27321907
MLKL octamer formation depends on alpha-helices 4 and 5. PMID: 27920255
The findings reported here indicate that palmitate induces RIP1/RIP3-dependent necrosis via MLKL-mediated pore formation of RAW 264.7 cells in the plasma membrane, which could provide a new mechanism to explain the link between elevated levels of free fatty acids (FFAs), palmitate in particular, and macrophage death. PMID: 27856241
The results indicate that RIP1 and MLKL contribute to necroptotic cell death after HCoV-OC43 infection to limit viral replication. PMID: 27795420
In AML, Mlkl expression is reduced. This leads to reduced activation of the inflammasome and reduced myeloid differentiation. PMID: 27411587
we demonstrate that the phosphorylation of Ser345 is not required for the interaction between RIPK3 and MLKL in the necrosome, but is essential for MLKL translocation, accumulation in the plasma membrane, and consequent necroptosis. PMID: 26024392
deficiency of RIPK3 or MLKL prevents oxalate crystal-induced acute kidney injury. PMID: 26817517
RIPK3 can promote NLRP3 inflammasome and IL-1beta inflammatory responses independent of MLKL and necroptotic cell death PMID: 25693118
Cisplatin stimulates RIP1/RP3/MLKL-dependent necrotic cell death in renal tubules, which finally causes renal dysfunction PMID: 25788533
These data reveal a potential role for RIPK3 as a suppressor of MLKL activation and indicate that phosphorylation can fine-tune the ability of MLKL to induce necroptosis. PMID: 26283547
MLKL acctivation unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death PMID: 25288762
Ectopic expression of ICP6, but not RHIM mutant ICP6, directly activated RIP3/MLKL-mediated necrosis. PMID: 25316792
Knockdown of RIPK3 or MLKL blocks TNF-induced necroptosis. PMID: 24434512
[review] Although studies have demonstrated that mixed lineage kinase domain-like (MLKL) protein is the only substrate of RIP3 kinase that is essential for necroptotic death, the molecular determinants acting downstream of MLKL remain ambiguous. PMID: 24556404
Data suggest that nucleotide- (ATP-) binding residues of human MLKL have divergently evolved from mouse Mlkl and conventional protein kinases; studies include small-angle X-ray scattering, thermal shift of nucleotide binding, and sequence alignment. PMID: 24219132
Neither Mlkl nor Rip3 deficiency provided protection against polymicrobial sepsis-induced animal death in the study of septic shock induced by CLP. PMID: 23835476
Toll-like receptor 3-mediated necrosis via TRIF, RIP3, and MLKL. PMID: 24019532
crystal structure determined; structure-guided mutation of the MLKL pseudoactive site resulted in constitutive, RIPK3-independent necroptosis, demonstrating that modification of MLKL is essential for propagation of the necroptosis pathway downstream of RIPK3 PMID: 24012422
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
Findings implicate MLKL as a key mediator of necrosis signaling downstream of the kinase RIP3. PMID: 22265413

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Subcellular Location

Cytoplasm. Cell membrane. Nucleus.

Protein Families

Protein kinase superfamily

Tissue Specificity

Highly expressed in thymus, colon, intestine, liver, spleen and lung. Expressed at much lower level in skeletal muscle, heart and kidney. Not detected in brain.

Database Links

KEGG: mmu:74568

STRING: 10090.ENSMUSP00000113718

UniGene: Mm.207971

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Code	CSB-YP861529MO
MSDS	MSDS
Size	$306
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Image	(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Recombinant Mouse Mixed lineage kinase domain-like protein (Mlkl)

Product Details

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