MYD88 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Western blot
Alternative names
MYD88D, MYD88_HUMAN, Mutant myeloid differentiation primary response 88, Myeloid differentiation marker 88, Myeloid differentiation primary response 88, Myeloid differentiation primary response gene, Myeloid differentiation primary response gene (88), Myeloid differentiation primary response gene 88, Myeloid differentiation primary response protein MyD88, OTTHUMP00000161718, OTTHUMP00000208595, OTTHUMP00000209058, OTTHUMP00000209059, OTTHUMP00000209060
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MYD88 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Western blot
- Disease
Carcinoma
- Concentration
Properties
- Gene name
MYD88
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 1 US: 1
- Adherent/suspension
Adherent
- Gender
Male
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- Culture medium
F-12K + 10% FBS
- Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (Human wild-type A549 cell line ab288558). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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.
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4 product images
Western blot - Human MYD88 knockout A549 cell line (ab286715)
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 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.
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 - Human MYD88 knockout A549 cell line (ab286715)
Western blot: Anti-MYD88 antibody (Anti-MyD88 antibody ab2064) 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 ab2064 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 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.
All lanes: Western blot - Anti-MyD88 antibody (Anti-MyD88 antibody ab2064) 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.
Observed band size: 35 kDa
Western blot - Human MYD88 knockout A549 cell line (ab286715)
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 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 (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
Western blot - Human MYD88 knockout A549 cell line (ab286715)
False colour image of Western blot: Anti-MyD88 antibody [EPR21824] 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 [EPR21824] ab219413 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 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. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-MyD88 antibody [EPR21824] (Anti-MyD88 antibody [EPR21824] ab219413) 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
SecondaryLanes 1 - 3: Goat anti-Mouse IgG H&L 680RD
Lanes 1 - 3: Goat anti-Rabbit IgG H&L 800CW
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 35 kDa
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