MICA KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout
Alternative names
FLJ36918, FLJ60820, HLA class I antigen, MGC111087, MGC21250, MHC class I chain related gene A protein, MHC class I chain related protein A, MHC class I chain related protein A HLA B HLA C, MHC class I polypeptide-related sequence A, MHC class I related protein, MICA_HUMAN, OTTHUMP00000029088, OTTHUMP00000044528, OTTHUMP00000165170, OTTHUMP00000165172, PERB11.1, Stress inducible class I homolog
Recommended products
MICA KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- MICA
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, 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 (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
MICA also known as MHC class I polypeptide-related sequence A is a protein encoded by the MICA gene. It has a molecular weight of approximately 62 kDa. MICA is a stress-induced ligand expressed on the surface of various cell types including epithelial cells fibroblasts and some tumor cells. This protein is part of the MHC class I chain-related (MIC) family which plays an important role in the immune system by interacting with natural killer (NK) cells and certain T cells.
- Biological function summary
The MICA protein acts as a ligand for the NKG2D receptor an activating receptor found on NK cells and some T-cell subsets. MICA does not require expression as part of a complex with β2-microglobulin for function. When cells express MICA they signal distress allowing immune cells to target and destroy potentially harmful cells such as those infected with viruses or transformed into cancer cells. This interaction is an important component of the immune surveillance system.
- Pathways
MICA functions within the natural cytotoxicity effector pathway heavily involved in immune responses. It is also relevant in the DNA damage response pathways. Within these pathways MICA interacts with proteins like NKG2D and plays a part in mediating cellular responses to stress and damage. These interactions enhance the capacity of immune cells to recognize and eliminate abnormal cells.
- Associated diseases and disorders
MICA is associated with conditions such as psoriasis and certain types of cancer. Studies have shown increased levels of soluble MICA in the sera of patients with psoriasis indicating a role in inflammation. Additionally in cancer MICA expression affects tumor immunity connecting to the NKG2D receptor to influence immune cell targeting and destruction of tumor cells. Researchers use tools such as MICA ELISA kits and imaging techniques to study its role in these diseases.
Product promise
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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.
2 product images
Western blot - Human MICA knockout A549 cell line (ab288893)
Western blot: Anti-MICA antibody [EPR22031] (Anti-MICA + MICB antibody [EPR22031] ab224702) 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 magenta. In Western blot, Anti-MICA + MICB antibody [EPR22031] ab224702 was shown to bind specifically to MICA. A band was observed at 43 kDa in wild-type A549 cell lysates with no signal observed at this size in MICA knockout cell line. To generate this image, wild-type and MICA 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-MICA + MICB antibody [EPR22031] (Anti-MICA + MICB antibody [EPR22031] ab224702) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: MICA knockout A549 cell lysate at 20 µg
Lane 3: U-2 OS cell lysate at 20 µg
Lane 4: Ramos cell lysate at 20 µg
Lane 5: MICA protein Recombinant human MICA protein (Active) ab182709 cell lysate at 0.2 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 43 kDa
Next Generation Sequencing - Human MICA knockout A549 cell line (ab288893)
94 bp deletion after Met 207 (allele 1); 2 bp deletion after Ala 228 (allele 2) of WT protein
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