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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| M | ICC, IP, WB | R | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABC604 |
| Description | Anti-MLKL Antibody, clone 3H1 |
| Alternate Names |
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| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified rat IgG in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mm NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-MLKL Antibody, clone 3H1 is validated for use in Immunocytochemistry, Immunoprecipitation, and Western Blotting for the detection of MLKL. |
| Key Applications |
|
| Application Notes | 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). |
| Biological Information | |
|---|---|
| Immunogen | 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). |
| Epitope | Brace region. |
| Clone | 3H1 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rat |
| Isotype | IgG |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein g purified. |
| UniProt Number |
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| Molecular Weight | ~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. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |