Recombinant
RabMAb

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

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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: 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

    • 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.

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

    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.

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
ICC/IF Use at an assay dependent concentration.
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

    • Immunocytochemistry/ Immunofluorescence - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)
      Immunocytochemistry/ Immunofluorescence - Anti-MLKL (phospho S345) antibody [EPR9515(2)] - BSA and Azide free (ab208910)This image is courtesy of an anonymous Abreview

      Ab196436 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. 

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

    • 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.

    References

    This product has been referenced in:

    • Cui YL  et al. Necroptosis as a potential therapeutic target in multiple organ dysfunction syndrome. Oncotarget 8:56980-56990 (2017). IHC-P ; Rat . Read more (PubMed: 28915647) »
    See 1 Publication for this product

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

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