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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MyD88 antibody [EPR590(N)] (ab133739)

  • Datasheet
  • SDS
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Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Immunocytochemistry/ Immunofluorescence - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • OI-RD Scanning - Anti-MyD88 antibody [EPR590(N)] (ab133739)
  • Anti-MyD88 antibody [EPR590(N)] (ab133739)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR590(N)] to MyD88
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-MyD88 antibody [EPR590(N)]
    See all MyD88 primary antibodies
  • Description

    Rabbit monoclonal [EPR590(N)] to MyD88
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to residues at the N-terminus of Human MyD88. (Q99836)

  • Positive control

    • Human kidney tissue. Jurkat, Molt-4, HepG2, K562, Raji cell lysates. Flow Cyt: HAP1-WT cells
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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

  • 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. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 1.16 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR590(N)
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Alternative Versions

    • Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)
    • HRP Anti-MyD88 antibody [EPR590(N)] (ab218705)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HepG2 cytoplasmic extract lysate (ab14659)
    • Jurkat cytoplasmic extract lysate (ab14842)
    • MOLT4 whole cell lysate (ab7912)
  • Recombinant Protein

    • Recombinant Human MyD88 protein (ab114217)

Applications

Our Abpromise guarantee covers the use of ab133739 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 1/500.
WB 1/1000 - 1/10000. Predicted molecular weight: 33 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified, use 1/50 - 1/100.

Flow Cyt 1/350.

For unpurified use 1/500 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Adapter protein involved in the Toll-like receptor and IL-1 receptor signaling pathway in the innate immune response. Acts via IRAK1, IRAK2, IRF7 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Increases IL-8 transcription. Involved in IL-18-mediated signaling pathway.
  • Tissue specificity

    Ubiquitous.
  • Involvement in disease

    Defects in MYD88 are the cause of MYD88 deficiency (MYD88D) [MIM:612260]; also known as recurrent pyogenic bacterial infections due to MYD88 deficiency. Patients suffer from autosomal recessive, life-threatening, often recurrent pyogenic bacterial infections, including invasive pneumococcal disease, and die between 1 and 11 months of age. Surviving patients are otherwise healthy, with normal resistance to other microbes, and their clinical status improved with age.
  • Sequence similarities

    Contains 1 death domain.
    Contains 1 TIR domain.
  • Domain

    The intermediate domain (ID) is required for the phosphorylation and activation of IRAK.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession Q99836 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4615 Human
    • Omim: 602170 Human
    • SwissProt: Q99836 Human
    • Unigene: 82116 Human
    • Alternative names

      • Mutant myeloid differentiation primary response 88 antibody
      • MYD 88 antibody
      • Myd88 antibody
      • MYD88_HUMAN antibody
      • MYD88D antibody
      • Myeloid differentiation marker 88 antibody
      • Myeloid differentiation primary response 88 antibody
      • Myeloid differentiation primary response gene (88) antibody
      • Myeloid differentiation primary response gene 88 antibody
      • Myeloid differentiation primary response gene antibody
      • Myeloid differentiation primary response protein MyD88 antibody
      • OTTHUMP00000161718 antibody
      • OTTHUMP00000208595 antibody
      • OTTHUMP00000209058 antibody
      • OTTHUMP00000209059 antibody
      • OTTHUMP00000209060 antibody
      see all

    Images

    • Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: MyD88 knockout HAP1 cell lysate (20 µg)
      Lane 3: Jurkat cell lysate (20 µg)
      Lane 4: MOLT4 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab133739 observed at 37 kDa. Red - loading control, ab7291, observed at 52 kDa.

      ab133739 was shown to specifically react with MyD88 when MyD88 knockout samples were used. Wild-type and MyD88 knockout samples were subjected to SDS-PAGE. ab133739 and ab7291 (loading control to alpha tubulin) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)

      Overlay histogram showing HAP1 wildtype (green line) and HAP1-MyD88 knockout cells (red line) stained with ab133739. The cells were fixed with 80% methanol (5 min)  and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab133739, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22°C.

      A rabbit IgG1 isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MyD88 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

      This antibody can also be used in HAP1 cells fixed with  4% formaldehyde (10 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

    • Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      All lanes : Anti-MyD88 antibody [EPR590(N)] (ab133739) at 1/5000 dilution (purified)

      Lane 1 : HepG2 cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : Jurkat cell lysate

      Lysates/proteins at 29 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 33 kDa
      Observed band size: 33 kDa



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Immunocytochemistry/ Immunofluorescence - Anti-MyD88 antibody [EPR590(N)] (ab133739)

      Immunofluorescence staining of Jurkat cells with purified ab133739 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab133739 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab133739 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Overlay histogram showing K562 cells fixed in 4% PFA and stained with purified ab133739 at a dilution of 1 in 350 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
    • Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Flow Cytometry - Anti-MyD88 antibody [EPR590(N)] (ab133739)

      Overlay histogram showing MCF7 cells stained with unpurified ab133739 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133739, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Western blot - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      All lanes : Anti-MyD88 antibody [EPR590(N)] (ab133739) at 1/1000 dilution (unpurified)

      Lane 1 : Jurkat cell lysate
      Lane 2 : Molt-4 cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : K562 cell lysate
      Lane 5 : Raji cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit at 1/2000 dilution

      Predicted band size: 33 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] (ab133739)

      Immunohistochemistry analysis of Myd88 expression in formalin-fixed, paraffin-embedded Human kidney tissue, using unpurified ab133739 at 1/50 dilution.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • OI-RD Scanning - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      OI-RD Scanning - Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-MyD88 antibody [EPR590(N)] (ab133739)
      Anti-MyD88 antibody [EPR590(N)] (ab133739)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (12)

    Publishing research using ab133739? Please let us know so that we can cite the reference in this datasheet.

    ab133739 has been referenced in 12 publications.

    • Yu X  et al. Asiatic acid ameliorates obesity-related osteoarthritis by inhibiting myeloid differentiation protein-2. Food Funct 11:5513-5524 (2020). PubMed: 32514515
    • Yuan J  et al. The suppressive role of miR-362-3p in epithelial ovarian cancer. Heliyon 6:e04258 (2020). PubMed: 32671239
    • Chen J  et al. Expression and Significance of MyD88 in Patients With Gastric Cardia Cancer in a High-Incidence Area of China. Front Oncol 10:559 (2020). PubMed: 32477927
    • Cui S  et al. Kaempferol protects lipopolysaccharide-induced inflammatory injury in human aortic endothelial cells (HAECs) by regulation of miR-203. Biomed Pharmacother 115:108888 (2019). PubMed: 31022599
    • Hou XX  et al. Aryl Hydrocarbon Receptor Modulates the Expression of TNF-a and IL-8 in Human Sebocytes via the MyD88-p65NF-?B/p38MAPK Signaling Pathways. J Innate Immun 11:41-51 (2019). PubMed: 30056444
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-MyD88 antibody [EPR590(N)]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (8)
    Sample
    Mouse Cell lysate - whole cell (Mouse bone marrow-derived macrophages)
    Specification
    Mouse bone marrow-derived macrophages
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

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    Verified customer

    Submitted Sep 02 2013

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