MLKL Antibody (YA289)

MLKL Antibody (YA289) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to MLKL.

MLKL, a pseudokinase, plays a pivotal role in TNF-induced necroptosis, a programmed cell death process characterized by plasma membrane damage and calcium influx. Despite lacking intrinsic protein kinase activity, MLKL is activated upon phosphorylation by RIPK3. This activation leads to homotrimerization, plasma membrane localization, and execution of necroptosis. Additionally, in response to orthomyxoviruses infection, MLKL can undergo nuclear necroptosis triggered by ZBP1 activation, resulting in disruption of the nuclear envelope and release of cellular DNA into the cytosol. MLKL's necroptotic function is facilitated by its binding to highly phosphorylated inositol phosphates, such as inositolhexakisphosphate (InsP6). This binding not only mediates the release of an N-terminal auto-inhibitory region but also requires RIPK3-dependent phosphorylation for full activation. Necrosulfonamide, a specific inhibitor of necroptosis, targets Cys-86 and effectively inhibits MLKL function.

Free

Q8NB16

Predicted band size: 54 kDa； Observed band size: 54 kDa

Protein A affinity purified.

Cytoplasm, Cell membrane, Golgi apparatus.

Non-conjugated

Unmodified

AB_3102361

Signal Transduction

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:1,000 ;IHC-P: 1:50

• 
  Western blot analysis of extracts from Hela (lane 2(20μg) , THP-1(lane 3(20μg) and U937(lane 4(20μg) using MLKL(HY-P80227) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using MLKL Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80227, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using MLKL Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80227, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-P

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human MLKL.AA range:333-471.