RecombinantRabMAb

Recombinant Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)

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Immunoprecipitation - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)
  • Dot Blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)
  • Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9515(2)] to MLKL (phospho S345) - BSA and Azide free
  • Suitable for: WB, IP, Dot blot
  • Reacts with: Mouse

Overview

  • Product name

    Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free
    See all MLKL primary antibodies

  • Description

    Rabbit monoclonal [EPR9515(2)] to MLKL (phospho S345) - BSA and Azide free

  • 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: 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

    • WB: L929 whole cell lysate treated with 20 ng/ml TNF alpha, 100 nM Smac mimetic, and 20 mM z-VAD for 8h and then harvested. IP: L929 whole cell lysate treated with TNF alpha+ Smac mimetic + z-VAD compound for 8h.

  • General notes

    ab208910 is the carrier-free version of ab196436 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab208910 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab208910 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
WB Use at an assay dependent concentration. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
IP Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.

Target

  • 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

  • Database links

    • Alternative names

      • 9130019I15Rik antibody
      • FLJ34389 antibody
      • hMLKL antibody
      • Mixed lineage kinase domain like antibody
      • Mixed lineage kinase domain like protein antibody
      • Mixed lineage kinase domain like pseudokinase antibody
      • Mixed lineage kinase domain-like protein antibody
      • Mlkl antibody
      • MLKL_HUMAN antibody
      see all

    Images

    • Immunoprecipitation - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)
      Immunoprecipitation - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)

      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 + 100 nM Smac mimetic + 20 µM z-VAD compound 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+ 100 nM Smac mimetic+ 20 µM z-VAD compound for 8h;10 µg (Input).
      Lane 2: ab196436 IP in L-929 whole cell lysate treated with 20 ng/ml TNF alpha+ 100 nM Smac mimetic+ 20 µM z-VAD compound for 8h.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab196436 in L-929 whole cell lysate treated with 20 ng/ml TNF alpha+ 100 nM Smac mimetic+ 20 µM z-VAD compound for 8h.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196436).

    • Dot Blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)
      Dot Blot - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)

      This Dot Blot data was generated using the same anti-MLKL (phospho S345) antibody clone, EPR9515(2), in a different buffer formulation (cat# 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/Dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)
      Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)

    References (3)

    Publishing research using ab208910? Please let us know so that we can cite the reference in this datasheet.

    ab208910 has been referenced in 3 publications.

    • Zhu Y  et al. Angiotensin II triggers RIPK3-MLKL-mediated necroptosis by activating the Fas/FasL signaling pathway in renal tubular cells. PLoS One 15:e0228385 (2020). PubMed: 32134954
    • Zhu Y  et al. AT1 and AT2 receptors modulate renal tubular cell necroptosis in angiotensin II-infused renal injury mice. Sci Rep 9:19450 (2019). PubMed: 31857626
    • Cui YL  et al. Necroptosis as a potential therapeutic target in multiple organ dysfunction syndrome. Oncotarget 8:56980-56990 (2017). IHC-P ; Rat . PubMed: 28915647

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