MABC604

Anti-MLKL Antibody, clone 3H1

clone 3H1, from rat

• Synonyma: Mixed lineage kinase domain-like protein, MLKL
• UNSPSC Code: 12352203
• NACRES: NA.41
• eCl@ss: 32160702
• Conjugate: unconjugated
• Clone: 3H1, monoclonal
• Application: ICC, IP, WB
• Citations: 123

Vlastnosti

• biological source: rat
• Quality Segment: 100
• conjugate: unconjugated
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 3H1, monoclonal
• species reactivity: mouse
• technique(s): immunocytochemistry: suitable, immunoprecipitation (IP): suitable, western blot: suitable
• isotype: IgG
• NCBI accession no.: NP_083281
• UniProt accession no.: Q9D2Y4
• shipped in: wet ice
• target post-translational modification: unmodified
• Gene Information: mouse ... Mlkl(74568)

Popis

General description

MLKL, also known as Mixed lineage kinase domain-like protein, and encoded by the gene MLKL, is an interesting protein that is required for the execution of programmed necrosis or necroptosis, for example in response to TNF-alpha induced necrosis. MLKL is a component of the "necrosome," the multiprotein complex that triggers tumor necrosis factor (TNF)-induced cell death by necroptosis. The programmed necrosis induced by TNF-α also requires the activities of the receptor-interacting serine-threonine kinases RIP1 and RIP3 and they too interact directly with the mixed lineage kinase domain-like protein MLKL. RIP3 is an essential upstream kinase in necroptosis. The pseudokinase MLKL functions as a substrate of RIP3 to mediate downstream signaling and when complexed to MLKL the RIP3-MLKL complex is stabilized by AMP-PNP to adopt an inactive conformation.

~52 kDa observed. 54.48/30.28 (human isoform 1/2) and 54.32/53.38 kDa (mouse isoform 1/2) calculated. Uncharacterized band(s) may be observed in some cell lysates.

Immunogen

Epitope: Brace region.

KLH-conjugated linear peptide corresponding to a sequence from the two-helix linker (Brace) region N-terminal to the pseudokinase domain of mouse MLKL (Hildebrand, J.M., et al. (2014). Proc. Natl. Acad. Sci. U.S.A. 111(42):15072-15077; Murphy, J.M., et al. (2013). Immunity. 39(3):443-453).

Application

Immunocytochemistry Analysis: A representative lot detected MLKL in mouse dermal fibroblasts (MDFs) from wild-type, but not Mlkl-/- mice (Courtesy of Prof. James M. Murphy, Walter and Eliza Hall Institute, Australia).

Immunoprecipitation Analysis: A representative lot immunoprecipitated MLKL from soluble extract of Mlkl-/- mouse dermal fibroblasts (MDFs) harboring doxycycline-inducible wild-type mouse MLKL expression construct only after, but not before doxycycline treatment (Murphy, J.M., et al. (2013). Immunity. 39(3):443-453).

Western Blotting Analysis: A representative lot detected Q-VD-OPh (TSQ; Cat. No. 551476) treatment-induced membrane translocation of murine and equine MLKL N-terminal fragment (a.a. 1-180 and 1-189, respectively) exogenously expressed in mouse dermal fibroblasts (MDFs) from Mlkl-/- mice (Tanzer, M.C., et al. (2016). Cell Death Differ.. In press).

Western Blotting Analysis: A representative lot detected MLKL in human HT-29 and U937 cells (Tanzer, M.C., et al. (2015). Biochem. J. 471(2):255-265).

Western Blotting Analysis: Representative lots detected MLKL membrane translocation in mouse dermal fibroblasts (MDFs) upon Q-VD-OPh (TSQ; Cat. No. 551476) treatment (Tanzer, M.C., et al. (2015). Biochem. J. 471(2):255-265; Hildebrand, J.M., et al. (2014). Proc. Natl. Acad. Sci. U.S.A. 111(42):15072-15077).

Western Blotting Analysis: Representative lots detected MLKL expression in L292 mouse fibroblasts, as well as in mouse dermal fibroblasts (MDFs), mouse embryonic fibroblasts (MEFs) and bone marrow derived macrophages (BMDMs) from wild-type, but not Mlkl-/- mice (Cook, W.D., et al. (2014). Cell Death Differ. 21(10):1600-1612; Murphy, J.M., et al. (2013). Immunity. 39(3):443-453).

Western Blotting Analysis: A representative lot detected recombinant full-length mouse MLKL as well as a.a. 1-180 and a.a. 124-464, but not a.a. 1-125 or a.a. 179-464, MLKL fragments (Hildebrand, J.M., et al. (2014). Proc. Natl. Acad. Sci. U.S.A. 111(42):15072-15077).

Western Blotting Analysis: A representative lot detected MLKL expression in all tissues tested except brain from wild-type mice, while no target band was seen in any tissues from Mlkl-/- mice (Murphy, J.M., et al. (2013). Immunity. 39(3):443-453).

This Anti-MLKL Antibody, clone 3H1 is validated for use in Immunocytochemistry, Immunoprecipitation, and Western Blotting for the detection of MLKL.

Physical form

Format: Purified

Purified rat IgG in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Analysis Note

Evaluated by Western Blotting in mouse heart tissue lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected MLKL in 10 µg of mouse heart tissue lysate.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Bezpečnostní informace

• Skladovací třída: 12 - Non Combustible Liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable