MABC1158

Anti-phospho-MLKL (Ser345) Antibody, clone 7C6.1

clone 7C6.1, from mouse

• Synonym(s): Mixed lineage kinase domain-like protein
• UNSPSC Code: 12352203
• NACRES: NA.43
• eCl@ss: 32160702
• Clone: 7C6.1, monoclonal
• Species reactivity: mouse
• Application: ELISA, ICC, WB
• Citations: 14

Properties

• biological source: mouse
• Quality Segment: 100
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 7C6.1, monoclonal
• species reactivity: mouse
• technique(s): ELISA: suitable, immunocytochemistry: suitable, western blot: suitable
• isotype: IgG2bκ
• NCBI accession no.: NP_001297542
• UniProt accession no.: Q9D2Y4
• shipped in: ambient
• target post-translational modification: phosphorylation (pSer345)
• Gene Information: mouse ... Mlkl(74568)

Description

General description

Mixed lineage kinase domain-like protein (UniProt: Q9D2Y4; also known as MLKL) is encoded by the Mlkl gene (Gene ID: 74568) in murine species. MLKL is a pseudokinase that plays a key role in TNF-induced necroptosis. MLKL is highly expressed in thymus, colon, intestine, liver, spleen and lung and is expressed at much lower level in skeletal muscle, heart and kidney. It is not detected in brain. MLKL is activated following phosphorylation by RIPK3 that induces a conformational switch, which is essential for necroptosis. The protein kinase domain of MLKL is catalytically inactive, but contains a pseudoactive site with an interaction between Lys-219 and Gln-343 residues. Upon phosphorylation by RIPK3, MLKL undergoes an active conformation and homotrimerization through its amino-terminal coiled-coil domain. Phosphorylation is also essential for homotrimerization of MLKL and its subsequent localization to the plasma membrane. Plasma membrane localization of MLKL is considered to be essential for Ca2+ influx, which is an early event of TNF-induced necroptosis. (Ref.: Cai, Z et al. (2014). Nat. Cell Biol. 16(1): 55-65).

~54 kDa observed; 54.32 kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Immunogen

KLH-conjugated linear peptide corresponding to 19 amino acids surrounding Ser345 from the C-terminal half of murine Mixed lineage kinase domain-like protein (MLKL).

Application

Detect phospho MLKL Ser345 using this mouse monoclonal Anti-phospho-MLKL (Ser345), clone 7C6.1, Cat. No. MABC1158. It has been tested in ELISA, Immunocytochemistry, and Western Blotting.

ELISA Analysis: A representative lot detected phospho-MLKL (Ser345) in ELISA applications (Rodriguez, D.A., et. al. (2016). 23(1):76-88).

Immunocytochemistry Analysis: A representative lot detected phospho-MLKL (Ser345) in Immunocytochemistry applications (Rodriguez, D.A., et. al. (2016). 23(1):76-88).

Western Blotting Analysis: A representative lot detected phospho-MLKL (Ser345) in Western Blotting applications (Rodriguez, D.A., et. al. (2016). 23(1):76-88).

Research Category
Apoptosis & Cancer

Biochem/physiol Actions

Clone 7C6.1 specifically detects MLKL phosphorylalated on Ser345. It targets a sequence of 19 amino acids surrounding Ser345 in the C-terminal region.

Physical form

Format: Purified

Protein G purified

Purified mouse monoclonal antibody IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Preparation Note

Stable for 1 year at 2-8°C from date of receipt.

Analysis Note

Evaluated by Western Blotting in MLKL -/- mouse embryonic fibroblasts expressing doxycycline-inducible wild-type MLKL and treated with TNF alpha and Z-VAD-FMK.

Western Blotting Analysis: A 1:4,000 dilution of this antibody detected phospho-MLKL Ser345 in 20 ug of lysate from MLKL -/- mouse embryonic fibroblasts expressing doxycycline-inducible wild-type MLKL-Flag-C construct and treated with TNF alpha (10 ng/mL) and Z-VAD-FMK (25 uM).

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable