Human TSPO (PBR) knockout HeLa cell line
- Advanced Validation
- What is this?
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TSPO KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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)
- WB
Lab
Western blot - Human TSPO (PBR) knockout HeLa cell line (AB264942)
Lanes 1- 2 : Merged signal (red and green). Green - ab109497 observed at 15 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
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 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. ab109497 and 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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/pbr-antibody-epr5384-ab109497'>ab109497</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human TSPO (PBR) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-tspo-pbr-knockout-hela-cell-lysate-ab257066'>ab257066</a>) at 20 µg
Predicted band size: 19 kDa
Observed band size: 17 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TSPO (PBR) knockout HeLa cell line (AB264942)
Allele-2 : 2 bp deletion in exon 2.
- Cell Culture
Unknown
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 seq
Unknown
Sanger Sequencing - Human TSPO (PBR) knockout HeLa cell line (AB264942)
Allele-1 : 5 bp deletion in exon 2.
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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).
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB109497
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PBR antibody [EPR5384] (AB109497)](https://content.abcam.com/products/images/pbr-antibody-epr5384-ab109497--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img454145.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com