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Knockout Tested Rabbit Recombinant Monoclonal MYD88 antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 5 publications.

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Images

Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (AB199247), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (AB199247), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (AB199247), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (AB199247), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (AB199247), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

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

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

Select an associated product type

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1 products for Alternative Version

Target data

Function

Adapter protein involved in the Toll-like receptor and IL-1 receptor signaling pathway in the innate immune response (PubMed:15361868, PubMed:18292575, PubMed:33718825). Acts via IRAK1, IRAK2, IRF7 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response (PubMed:15361868, PubMed:24316379, PubMed:19506249). Increases IL-8 transcription (PubMed:9013863). Involved in IL-18-mediated signaling pathway. Activates IRF1 resulting in its rapid migration into the nucleus to mediate an efficient induction of IFN-beta, NOS2/INOS, and IL12A genes. Upon TLR8 activation by GU-rich single-stranded RNA (GU-rich RNA) derived from viruses such as SARS-CoV-2, SARS-CoV and HIV-1, induces IL1B release through NLRP3 inflammasome activation (PubMed:33718825). MyD88-mediated signaling in intestinal epithelial cells is crucial for maintenance of gut homeostasis and controls the expression of the antimicrobial lectin REG3G in the small intestine (By similarity).

Alternative names

Myeloid differentiation primary response protein MyD88, MYD88

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal MYD88 antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 5 publications.

Alternative names

Myeloid differentiation primary response protein MyD88, MYD88

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR590(N)

Purification technique

Affinity purification Protein A

Dissociation constant

1.16 x 10-10 M

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab199247 is the carrier-free version of Anti-MyD88 antibody [EPR590(N)] ab133739.

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

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MyD88 standing for myeloid differentiation primary response 88 is a cytoplasmic adaptor protein with a molecular weight of approximately 33 kDa. This protein is expressed widely in immune cells including monocytes macrophages and dendritic cells. MyD88 serves a major role in the signaling pathways for the innate immune system. It acts as a linker transmitting signals from toll-like receptors (TLRs) and interleukin-1 receptors (IL-1Rs) to downstream signaling molecules ultimately activating transcription factors.

Biological function summary

MyD88 plays a significant role in mediating immune responses by forming part of a complex that includes IRAK kinases and TRAF6. When TLRs or IL-1Rs activate MyD88 this adaptor protein recruits IRAK4 which then phosphorylates IRAK1 or IRAK2. This cascade promotes the activation of NF-κB and MAPK pathways leading to the production of inflammatory cytokines. The MyD88-dependent pathway is integral to innate immunity influencing how the body responds to pathogen infection and inflammation.

Pathways

MyD88 integrates into both the TLR signaling and IL-1R signaling pathways. Key related proteins in these pathways include interleukin-1 receptor-associated kinase (IRAK) and tumor necrosis factor receptor-associated factor 6 (TRAF6). MyD88 initiates the recruitment and activation of IRAK1 and IRAK4 following receptor engagement leading to subsequent activation of downstream signals. As part of these pathways MyD88 mediates cellular responses important for immune system signaling and inflammatory response regulation.

Associated diseases and disorders

MyD88 involvement is notable in the context of oncological and autoimmune diseases. Mutations or dysregulation of MyD88 are implicated in conditions like lymphoma and rheumatoid arthritis. In lymphoma the mutation usually results in constant activation of NF-κB leading to unchecked cell growth. As for rheumatoid arthritis an overactive immune response due to MyD88 can cause additional inflammation affecting joint tissues. In these conditions MyD88 interaction with IRAK4 is significant given their mutual role in immune and inflammatory pathways.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-MyD88 knockout cells (red line) stained with Anti-MyD88 antibody [EPR590(N)] 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 (Anti-MyD88 antibody [EPR590(N)] ab133739, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG1 isotype control antibody  (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

  • Immunocytochemistry/ Immunofluorescence - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Immunofluorescence staining of Jurkat cells with purified Anti-MyD88 antibody [EPR590(N)] ab133739 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 Anti-MyD88 antibody [EPR590(N)] ab133739 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Immunohistochemical staining of paraffin embedded human cerebral cortex with purified Anti-MyD88 antibody [EPR590(N)] ab133739 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) 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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

  • Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Overlay histogram showing K562 cells fixed in 4% PFA and stained with purified Anti-MyD88 antibody [EPR590(N)] 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). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

  • Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Overlay histogram showing MCF7 cells stained with unpurified Anti-MyD88 antibody [EPR590(N)] 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 (Anti-MyD88 antibody [EPR590(N)] ab133739, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Immunohistochemistry analysis of Myd88 expression in formalin-fixed, paraffin-embedded Human kidney tissue, using unpurified Anti-MyD88 antibody [EPR590(N)] ab133739 at 1/50 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

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

  • OI-RD Scanning - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    OI-RD Scanning - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

  • Western blot - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Western blot - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    False colour image of Western blot: Anti-MyD88 antibody [EPR590(N)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-MyD88 antibody [EPR590(N)] ab133739 was shown to bind specifically to MyD88. A band was observed at 35 kDa in wild-type A549 cell lysates with no signal observed at this size in MYD88 knockout cell line Human MYD88 knockout A549 cell line ab286715 (knockout cell lysate ab290793). To generate this image, wild-type and MYD88 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

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

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: MYD88 knockout A549 cell lysate at 20 µg

    Lane 3: HEK-293 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 33 kDa

    Observed band size: 35 kDa

  • Western blot - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247), expandable thumbnail

    Western blot - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MyD88 antibody [EPR590(N)] ab133739).

    Western blot: Anti-MYD88 antibody [EPR590(N)] (Anti-MyD88 antibody [EPR590(N)] ab133739) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-MyD88 antibody [EPR590(N)] ab133739 was shown to bind specifically to MYD88. A band was observed at 35 kDa in wild-type A549 cell lysates with no signal observed at this size in MYD88 knockout cell line Human MYD88 knockout A549 cell line ab286715 (knockout cell lysate ab290793). To generate this image, wild-type and MYD88 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    Lanes 1 - 3: Western blot - Anti-MyD88 antibody [EPR590(N)] (Anti-MyD88 antibody [EPR590(N)] ab133739) at 1/1000 dilution

    Lanes 1 - 3: Western blot - Anti-MyD88 antibody [EPR590(N)] - BSA and Azide free (ab199247)

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: MYD88 knockout A549 cell lysate at 20 µg

    Lane 3: HEK-293 cell lysate at 20 µg

    Performed under reducing conditions.

    Observed band size: 35 kDa

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Product protocols

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