MABC604
Anti-MLKL Antibody, clone 3H1
clone 3H1, from rat
Synonym(s):
Mixed lineage kinase domain-like protein, MLKL
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
biological source
rat
Quality Level
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.
UniProt accession no.
shipped in
wet ice
target post-translational modification
unmodified
Gene Information
mouse ... Mlkl(74568)
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.
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).
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.
Quality
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.
Western Blotting Analysis: 0.5 µg/mL of this antibody detected MLKL in 10 µg of mouse heart tissue lysate.
Target description
~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.
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.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
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Flash Point(F)
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Flash Point(C)
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Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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D M Moujalled et al.
Cell death & disease, 5, e1086-e1086 (2014-03-01)
Necroptosis is a mechanism by which cells can kill themselves that does not require caspase activity or the presence of the pro-apoptotic Bcl-2 family members Bax or Bak. It has been reported that RIPK3 (receptor interacting protein kinase 3) activates
RIPK1- and RIPK3-induced cell death mode is determined by target availability.
Cook, WD; Moujalled, DM; Ralph, TJ; Lock, P; Young, SN; Murphy, JM; Vaux, DL
Cell Death and Differentiation null
M C Tanzer et al.
Cell death and differentiation, 23(7), 1185-1197 (2016-02-13)
The pseudokinase, MLKL (mixed-lineage kinase domain-like), is the most terminal obligatory component of the necroptosis cell death pathway known. Phosphorylation of the MLKL pseudokinase domain by the protein kinase, receptor interacting protein kinase-3 (RIPK3), is known to be the key
A V Jacobsen et al.
Cell death & disease, 7, e2051-e2051 (2016-01-19)
Necroptosis is a caspase-independent form of regulated cell death that has been implicated in the development of a range of inflammatory, autoimmune and neurodegenerative diseases. The pseudokinase, Mixed Lineage Kinase Domain-Like (MLKL), is the most terminal known obligatory effector in
X M Zhao et al.
Cell death & disease, 7, e2089-e2089 (2016-02-13)
The pseudokinase mixed lineage kinase domain-like protein (MLKL) is a key component of tumor necrosis factor (TNF)-induced necroptosis and plays a crucial role in necroptosis execution. However, the mechanisms that control MLKL activity are not completely understood. Here, we identify
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