Abcam, ab263892, Human MLKL ELISA Kit

Size: 1 x 96Tests
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
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)

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.

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

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.