TAP2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 6 and 1 bp insertion in exon 6.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 6 and 1 bp insertion in exon 6
Alternative names
ABC transporter, MHC 2, ABC18, ABCB3, APT-2, ATP binding cassette, sub family B (MDR/TAP), member 3, ATP-binding cassette sub-family B member 3, Antigen peptide transporter 2, D6S217E, Peptide supply factor 2, Peptide transporter PSF2, Peptide transporter TAP2, Peptide transporter involved in antigen processing 2, RING 11, Really interesting new gene 11 protein, TAP2_HUMAN, Transporter 2 ATP binding cassette sub family B, Transporter 2, ABC (ATP binding cassette, Transporter 2, ATP binding cassette, sub family B (MDR/TAP)
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TAP2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 6 and 1 bp insertion in exon 6.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 6 and 1 bp insertion in exon 6
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- TAP2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
- Culture medium
- DMEM (High Glucose) + 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
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
TAP2 also known as Transporter 2 ATP Binding Cassette Subfamily B Member has a mass of approximately 76 kDa. It functions as a part of the peptide-loading complex where it plays a mechanical role in transporting antigen peptides into the endoplasmic reticulum. TAP2 is expressed primarily in antigen-presenting cells such as B cells dendritic cells and macrophages. It is an integral membrane protein located in the membrane of the endoplasmic reticulum.
- Biological function summary
The TAP2 protein operates as an important component of the immune system by facilitating the processing and presentation of antigens. It partners with TAP1 and forms a heterodimer complex essential for the transportation of peptides that are 8-16 amino acids long. This action is fundamental for the proper presentation of antigens on major histocompatibility complex (MHC) class I molecules which are pivotal in the recognition of infected or abnormal cells by cytotoxic T lymphocytes.
- Pathways
The TAP2 protein plays a significant role in the antigen processing and presentation pathways. It directly impacts the MHC class I pathway by aiding in the selection and transport of peptides that bind to MHC class I molecules. TAP2 functions alongside related proteins such as TAP1 and tapasin which help facilitate the loading of these peptides onto MHC class I molecules in the endoplasmic reticulum preparing them for cell surface presentation.
- Associated diseases and disorders
Mutations or deficiencies in the TAP2 gene are associated with bare lymphocyte syndrome type 1 (BLS1) a rare immunodeficiency disorder. This syndrome relates to a shortage of antigen presentation on MHC class I molecules leading to increased susceptibility to viral infections. TAP2 along with its partner protein TAP1 is critical in maintaining regular immune surveillance and any disruption in their function can result in profound immunological consequences including impaired cytotoxic T cell responses.
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3 product images
Western blot - Human TAP2 knockout HeLa cell line (ab265426)
False colour image of Western blot: Anti-TAP2 antibody staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-TAP2 antibody ab180611 was shown to bind specifically to TAP2. A band was observed at 75 kDa in wild-type HeLa cell lysates with no signal observed at this size in Tap2 knockout cell line ab265426 (knockout cell lysate Human TAP2 knockout HeLa cell lysate ab258712). To generate this image, wild-type and Tap2 knockout HeLa 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-TAP2 antibody (Anti-TAP2 antibody ab180611) at 1/500 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Tap2 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human TAP2 knockout HeLa cell line (ab265426)
Performed under reducing conditions.
Predicted band size: 76 kDa
Observed band size: 75 kDa
Sanger Sequencing - Human TAP2 knockout HeLa cell line (ab265426)
Allele-1: 16 bp deletion in exon 6.
Sanger Sequencing - Human TAP2 knockout HeLa cell line (ab265426)
Allele-2: 1 bp insertion in exon 6.
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