Recombinant Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)
Overview
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Product name
Anti-MLKL (phospho S345) antibody [EPR9515(2)]
See all MLKL primary antibodies -
Description
Rabbit monoclonal [EPR9515(2)] to MLKL (phospho S345) -
Host species
Rabbit -
Specificity
MLKL pS345 is a trigger for necroptosis. It is only detectable in infection/cellular damaged (PMID:29229989) or aging tissue (PMID: 28807105) but not in normal tissues. -
Tested applications
Suitable for: ICC/IF, WB, IP, Dot blotmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Mouse MLKL aa 300-400 (phospho S345). The exact sequence is proprietary.
Database link: Q9D2Y4 -
Positive control
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General notes
This antibody was developed through collaboration with the lab of Xiaodong Wang at the National Institute of Biological Sciences, Beijing.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR9515(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab196436 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | 1/100. | |
| WB | 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). | |
| IP | 1/150. | |
| Dot blot | 1/1000. |
Target
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Sequence similarities
Belongs to the protein kinase superfamily.
Contains 1 protein kinase domain. -
Domain
The protein kinase domain is predicted to be catalytically inactive. - Information by UniProt
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Database links
- Entrez Gene: 74568 Mouse
- SwissProt: Q9D2Y4 Mouse
- Unigene: 207971 Mouse
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Alternative names
- 9130019I15Rik antibody
- FLJ34389 antibody
- hMLKL antibody
see all
Images
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![Western blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)](https://www.abcam.com/ps/products/196/ab196436/Images/ab196436-284782-anti-mlkl-phospho-s345-antibody-epr95152-western-blot.jpg)
Western blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)All lanes : Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436) at 1/1000 dilution
Lane 1 : Untreated L-929 (Mouse connective tissue fibroblast cells) whole cell lysate
Lane 2 : L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642), 100 nM Smac mimetic, and 20 µM z-VAD (ab120382) for 8 h and then harvested.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)](https://www.abcam.com/ps/products/196/ab196436/Images/ab196436-355295-anti-mlkl-antibody-immunocytochemistry-rat-cells.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)This image is courtesy of an annonymous AbreviewParaformaldehyde-fixed, triton X-100 permeabilized Rat Cell (primary hippocampal neurons treated with TNF-a/ZVA) labeling MLKL (phospho S345) (Green) using ab196436 at 1/100 dilution in ICC/IF analysis. A Invitrogen A11034 Goat polyclonal Alexa Fluor® 488 was used as a secondary antibody.
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![Immunoprecipitation - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)](https://www.abcam.com/ps/products/196/ab196436/Images/ab196436-242136-anti-mlkl-phospho-s345-antibody-epr95152-immunoprecipitation.jpg)
Immunoprecipitation - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)MLKL (phospho S345) was immunoprecipitated from 1mg of L-929 (Mouse connective tissue fibroblast cells) whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic + 20 µM z-VAD compound (ab120382) for 8h using ab196436 at 1/150 dilution. Western blot was performed from the immunoprecipitate using ab196436 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h;10 µg (Input).
Lane 2: ab196436 IP in L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab196436 in L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)](https://www.abcam.com/ps/products/196/ab196436/Images/ab196436-308647-anti-mlkl-phospho-s345-antibody-epr95152-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)This image is courtesy of an anonymous AbreviewAb196436 staining MLKL in mouse cardiomyocytes by Immunocytochemistry. Samples were fixed with paraformaldehyde, permeabilized with 0.03% Triton x-100 and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody at 1/100 dilution for 12 hours. An Alexa Fluor® anti- rabbit was used as a secondary antibody at 1/200 dilution.
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![Western blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)](https://www.abcam.com/ps/products/196/ab196436/Images/ab196436-313529-anti-mlkl-phospho-s345-antibody-epr95152-western-blot.jpg)
Western blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)All lanes : Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436) at 1/1000 dilution
Lane 1 : L-929 treated with 20 ng/ml TNF alpha (ab9642), 100 nM Smac mimetic, and 20 µM z-VAD (ab120382) for 8 h, whole cell lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse colon tissue lysate
Lane 4 : Mouse lung tissue lysate
Lane 5 : Mouse retina tissue lysate
Lane 6 : Mouse liver tissue lysate
Lane 7 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: 50 secondsBlocking and diluting buffer: 5% NFDM/TBST.
MLKL pS345 is a trigger for necroptosis. It is only detectable in infection/cellular damaged (PMID:29229989) or aging tissue (PMID: 28807105) but not in normal tissues. -
![Dot Blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)](https://www.abcam.com/ps/products/196/ab196436/Images/ab196436-242138-anti-mlkl-phospho-s345-antibody-epr95152-dot-blot.jpg)
Dot Blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436)Dot blot analysis of MLKL (phospho S345) peptide (Lane 1), and non-phospho peptide (Lane 2), labeled using ab196436 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
References
ab196436 has been referenced in 78 publications.
- Patel S et al. RIP1 inhibition blocks inflammatory diseases but not tumor growth or metastases. Cell Death Differ 27:161-175 (2020). PubMed: 31101885
- Kadigamuwa C et al. Role of Retinoic Acid Receptor-? in DNA Damage-Induced Necroptosis. iScience 17:74-86 (2019). PubMed: 31255985
- Qiu Y et al. Shifting the balance of autophagy and proteasome activation reduces proteotoxic cell death: a novel therapeutic approach for restoring photoreceptor homeostasis. Cell Death Dis 10:547 (2019). PubMed: 31320609
- Lim J et al. Autophagy regulates inflammatory programmed cell death via turnover of RHIM-domain proteins. Elife 8:N/A (2019). PubMed: 31287416
- Lu J et al. Melatonin Suppresses Microglial Necroptosis by Regulating Deubiquitinating Enzyme A20 After Intracerebral Hemorrhage. Front Immunol 10:1360 (2019). PubMed: 31258534
