TSPO KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2.
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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, 2 bp deletion in exon 2 and 5 bp deletion in exon 2
Alternative names
BPBS, BZRP, Benzodiazapine receptor (peripheral), Benzodiazepine peripheral binding site, IBP, Isoquinoline carboxamide-binding protein, MBR, Mitochondrial benzodiazepine receptor, PKBS, PTBR, PTBZR02, Peripheral benzodiazepine receptor, Peripheral benzodiazepine receptor-related protein, Peripheral-type benzodiazepine receptor, Ptbzr, RATPTBZR02, TSPOA_HUMAN, Translocator protein, Tspo1, mDRC, pk18, translocator protein (18kDa)
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TSPO KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2.
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, 2 bp deletion in exon 2 and 5 bp deletion in exon 2
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- TSPO
- 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
Recommended control: Human wild-type HeLa cell line (ab255448). 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
PBR also known as the Peripheral Benzodiazepine Receptor is a protein predominantly found in the outer mitochondrial membrane. It has alternate names like Translocator Protein (TSPO) and the mass of this protein is approximately 18 kDa. PBR is expressed in various tissues but shows high levels in steroidogenic tissues like adrenal glands as well as in the brain heart liver and kidneys. The abundant presence in these tissues highlights its importance in a variety of physiological functions.
- Biological function summary
PBR interacts with cholesterol for the synthesis of steroid hormones making it important for steroidogenesis. PBR is a part of the Mitochondrial Permeability Transition Pore complex (MPTP) involved in regulating the transport of molecules across the mitochondrial membrane. Through its association with the MPTP PBR plays a significant role in mitochondrial functions such as apoptosis and energy metabolism. The interaction with other molecules also includes the binding with benzodiazepines impacting processes like immune response and cell proliferation.
- Pathways
PBR plays a role in the steroid biosynthesis and apoptosis pathways. It interfaces with the StAR (Steroidogenic Acute Regulatory) protein to facilitate cholesterol transport into mitochondria the initial step in steroid hormone production. PBR is also involved in pathways that regulate apoptosis and mitochondrial function linking it to different cellular processes through interactions with proteins like VDAC (Voltage-Dependent Anion Channel).
- Associated diseases and disorders
PBR has relevance to conditions such as neurodegenerative diseases and cancer. PBR expression changes in disorders like Alzheimer's disease where it might reflect mitochondrial dysfunctions. It has also been associated with certain cancers where aberrant PBR activity might contribute to altered cell proliferation and apoptosis. Proteins like caspases are involved in the apoptotic pathways connected with PBR highlighting its involvement in disease processes.
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4 product images
Western blot - Human TSPO (PBR) knockout HeLa cell line (ab264942)
Lanes 1- 2: Merged signal (red and green). Green - Anti-PBR antibody [EPR5384] ab109497 observed at 15 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.Anti-PBR antibody [EPR5384] ab109497 was shown to react with PBR in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264942 (knockout cell lysate Human TSPO (PBR) knockout HeLa cell lysate ab257066) was used. Wild-type HeLa and TSPO knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-PBR antibody [EPR5384] ab109497 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-PBR antibody [EPR5384] (Anti-PBR antibody [EPR5384] ab109497) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: TSPO (PBR) knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human TSPO (PBR) knockout HeLa cell line (ab264942)
Performed under reducing conditions.
Predicted band size: 112 kDa, 13 kDa, 134 kDa, 152 kDa, 17 kDa, 19 kDa, 25 kDa, 26 kDa, 28 kDa, 29 kDa, 37 kDa, 38 kDa, 39 kDa, 42 kDa, 43 kDa, 49 kDa, 50 kDa, 52 kDa, 56 kDa, 58 kDa, 68 kDa, 69 kDa, 71 kDa, 72 kDa, 74 kDa, 80 kDa, 81 kDa, 83 kDa, 87 kDa
Observed band size: 112 kDa, 15 kDa, 36 kDa, 38 kDa, 42 kDa, 45 kDa, 55 kDa, 60 kDa, 74 kDa, 80 kDa, 87 kDa
Cell Culture - Human TSPO (PBR) knockout HeLa cell line (ab264942)
Representative images of TSPO knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human TSPO (PBR) knockout HeLa cell line (ab264942)
Allele-2: 2 bp deletion in exon 2.
Sanger Sequencing - Human TSPO (PBR) knockout HeLa cell line (ab264942)
Allele-1: 5 bp deletion in exon 2.
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