AB263892

Human MLKL ELISA Kit

Recombinant
SimpleStep
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Human MLKL ELISA Kit is a single-wash 90-min Simplestep used to quantify Human MLKL with a sensitivity of 6.29 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

Mixed lineage kinase domain-like protein, hMLKL, MLKL

3 Images

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

sELISA

Supplier Data

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

Representative image of Human MLKL ELISA Kit (ab263892)

Components shown from left to right :

- 10X Wash Buffer PT (ab206977)

- 10X Human MLKL Capture Antibody

- 10X Human MLKL Detector Antibody

- Human MLKL Lyophilised Purified Protein (2 vials)

- 50X Cell Extraction Enhancer Solution (ab193971)

- Sample diluent NS

- TMB solution

- Stop Solution

- 5X Cell Extraction Buffer PTR (ab193970)

- Antibody diluent 5BI

SimpleStep Pre-Coated 96-Well Microplate (ab206978) - shown behind bottles / vials.

Plate Seals - shown underneath bottles / vials.

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the ELISA kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit

sELISA

Supplier Data

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

Interpolated concentrations of native MLKL in Human cell culture extract based on a 75 μg/mL extract load.

The concentrations of MLKL were measured in duplicate and interpolated from the MLKL standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MLKL concentration was determined to be 741.83 pg/mL in HUVEC cell extract.

sELISA

Unknown

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

Example of Human MLKL standard curve in 1X Cell Extraction Buffer PTR.

The MLKL standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 6.29 pg/mL

Range

78.13 - 5000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Human MLKL ELISA Kit (ab263892) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of MLKL protein in cell lysate. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human MLKL with 6.29 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Mixed lineage kinase domain-like (MLKL) is a pseudokinase containing a protein kinase-like domain that plays a critical role in tumor necrosis factor (TNF)-induced necroptosis as a functional substrate of receptor-interacting protein kinase 3 (RIP3). MLKL, together with receptor-interacting protein 1 (RIP1) and RIP3, form a necrosis signaling complex termed the necrosome, within which MLKL is phosphorylated at T357 and S358 residues by RIP31. Once phosphorylated, MLKL oligomers localize to the plasma membrane and form disrupting pores, capable of regulating the influx of Na+ and Ca2+ rupturing the cell membrane. Necroptosis requires MLKL and RIP3 and is implicated in the pathogenesis of various diseases through its role in necroptosis. Human MLKL shares 69% and 68% sequence homology with mouse and rat, respectively.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "6.8" }, { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "10.2" } ]

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Recovery

[ { "sample": "Cell Lysate", "range": "82 - 100 %", "average": "= 90" } ]

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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab263892-1x96Tests", "assetComponentDetails": [ { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB263892-CMP05", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB263892-CMP12", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB263892-CMP07", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB263892-CMP13", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB263892-CMP06", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB263892-CMP11", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human MLKL Lyophilized Recombinant Protein", "number":"AB263892-CMP03", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human MLKL Capture Antibody", "number":"AB263892-CMP09", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 5BI", "number":"AB263892-CMP02", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB263892-CMP08", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human MLKL Detector Antibody", "number":"AB263892-CMP10", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB263892-CMP04", "productcode":"" } ] } } }

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Properties and storage information

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

MLKL also known as mixed lineage kinase domain-like protein plays a critical role in the process of necroptosis a form of programmed cell death. The MLKL protein has a molecular weight of approximately 54 kDa. The protein exists mainly within the cytoplasm but translocates to the plasma membrane during cell death execution. Expression of MLKL happens in various tissues indicating its wide biological importance. Phosphorylation of MLKL often referred to as p-MLKL is key to triggering its activity marking the transition from an inactive to an active state during necroptosis.

Biological function summary

The MLKL protein acts as an executioner of cell death by forming a complex that disrupts the plasma membrane integrity. This process is downstream of receptor-interacting serine/threonine-protein kinase 3 (RIPK3) which phosphorylates MLKL to form the active necrosome complex. Active MLKL oligomerizes and migrates towards the inner leaflet of the plasma membrane binding to phosphatidylinositol phosphates which assists in pore formation and cellular rupture. The ability to measure MLKL activity levels such as via MLKL ELISA kits is important for understanding necrotic processes in detailed studies.

Pathways

MLKL is integrally involved in the necroptotic pathway alongside RIPK1 and RIPK3 which are key initiators of necroptosis. Phosphorylated MLKL acts downstream of RIPK3 resulting in cell death without caspase activation distinguishing necroptosis from apoptosis. MLKL and RIPK3 are tightly linked within this pathway with MLKL phosphorylation serving as a vital event for the execution phase. The necroptosis pathway is part of larger networks including inflammatory response pathways highlighting the importance of MLKL's role beyond sheer cell death.

MLKL has been implicated in various inflammatory conditions and neurodegenerative diseases. The dysregulation of necroptosis can contribute to disorders such as inflammatory bowel disease and amyotrophic lateral sclerosis. In inflammatory bowel disease increased levels of p-MLKL might lead to excessive cell death exacerbating inflammation. Similarly in neurodegenerative disorders the harmful activation of MLKL may accelerate neuronal cell death. Key interactions with proteins like RIPK3 and RIPK1 highlight MLKL's involvement in these pathological processes making it a potential target for therapeutic intervention.

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Product protocols

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Target data

Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process (PubMed : 22265413, PubMed : 22265414, PubMed : 22421439, PubMed : 24316671). Does not have protein kinase activity (PubMed : 22265413, PubMed : 22265414, PubMed : 22421439, PubMed : 24316671). 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 (PubMed : 22265413, PubMed : 22265414, PubMed : 22421439, PubMed : 24316671). In addition to TNF-induced necroptosis, necroptosis can also take place in the nucleus in response to orthomyxoviruses infection : following activation by ZBP1, MLKL is phosphorylated by RIPK3 in the nucleus, triggering disruption of the nuclear envelope and leakage of cellular DNA into the cytosol.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 (By similarity). Binds to highly phosphorylated inositol phosphates such as inositolhexakisphosphate (InsP6) which is essential for its necroptotic function (PubMed : 29883610).

See full target information MLKL

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Product promise

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Human MLKL ELISA Kit

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

Sandwich ELISA - Human MLKL ELISA Kit (AB263892)

Key facts

Detection method

Sample types

Reacts with

Assay type

Sensitivity

Range

Assay time

Assay Platform

Reactivity data

Product details

Precision

Recovery

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

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Product promise