AB279862

Phospho-MLKL (S345) ELISA Kit

Name: Phospho-MLKL (S345) ELISA Kit (ab279862) | Abcam
Brand: Abcam Limited
SKU: AB279862
Price: 1035 USD
Availability: InStock

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Phospho-MLKL (S345) ELISA Kit is a Semi-quantitative ELISA for the measurement of Phospho-MLKL (S345) in Human in Cell/Tissue Extracts samples.

View Alternative Names

Mixed lineage kinase domain-like protein, hMLKL, MLKL

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Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

sELISA

Supplier Data

Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

Positive Control.

HT29 cells were treated with Calyculin A.

Solubilize cells at 4 x 10⁷ cells/ml in Cell Lysate Buffer.

Serial dilutions of lysates were analyzed in this ELISA.

sELISA

Supplier Data

Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

HT29 cells treated with Calyculin A. HT29 cells were treated with Calyculin A.

sELISA

Supplier Data

Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

HT29 cells treated with Calyculin A. HT29 cells were treated with Calyculin A.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Semi-quantitative

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": { "ELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Phospho-MLKL (S345) ELISA Kit (ab279862) is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated MLKL protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-MLKL. An anti-pan MLKL antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and MLKL present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-MLKL (S345) antibody is used to detect phosphorylated MLKL. After washing away unbound antibody, HRP conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of MLKL (S345) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

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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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab279862-1x96Tests", "assetComponentDetails": [ { "size":"1 x 25 µL", "name":"HRP-conjugated anti-rabbit IgG concentrate (500X)", "number":"AB279862-CMP01", "productcode":"" }, { "size":"1 x 5 mL", "name":"2X Cell lysate buffer", "number":"AB279862-CMP09", "productcode":"" }, { "size":"1 x 25 mL", "name":"20X Wash Buffer", "number":"AB279862-CMP03", "productcode":"" }, { "size":"1 x 8 mL", "name":"Stop Solution", "number":"AB279862-CMP07", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Rabbit anti-phospho-MLKL (S345)-antibody", "number":"AB279862-CMP06", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Pan-MLKL Coated Microplate", "number":"AB279862-CMP04", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Positive Control - treated HT29 cell lysate", "number":"AB279862-CMP02", "productcode":"" }, { "size":"1 x 15 mL", "name":"5X Assay Diluent", "number":"AB279862-CMP05", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB One-Step Substrate Reagent", "number":"AB279862-CMP08", "productcode":"" } ] } } }

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Properties and storage information

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°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 phospho S345

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Product promise

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Phospho-MLKL (S345) ELISA Kit

Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

Sandwich ELISA - Phospho-MLKL (S345) ELISA Kit (AB279862)

Key facts

Detection method

Sample types

Reacts with

Assay type

Assay Platform

Reactivity data

Product details

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

Complementary Products

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Primary Antibodies

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