Product nameHuman TSPO (PBR) knockout HeLa cell lysate
See all PBR kits
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.
*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab260122 - Human TSPO knockout HeLa cell lysate 1 x 100µg ab255552 - Human wild-type HeLa cell lysate 1 x 100µg
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
STR AnalysisAmelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
FunctionResponsible for the manifestation of peripheral-type benzodiazepine recognition sites and is most likely to comprise binding domains for benzodiazepines and isoquinoline carboxamides. May play a role in the transport of porphyrins and heme. Plays a role in the transport of cholesterol across mitochondrial membranes in steroidogenic cells.
Tissue specificityFound in many tissue types. Expressed at the highest levels under normal conditions in tissues that synthesize steroids.
Sequence similaritiesBelongs to the TspO/BZRP family.
Cellular localizationMitochondrion membrane.
- Information by UniProt
- Benzodiazapine receptor (peripheral)
- Benzodiazepine peripheral binding site
KO cell lines
KO cell pellets
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab257066 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: TSPO knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab109497 observed at 15 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109497 Anti-PBR antibody [EPR5384] was shown to specifically 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 and PBR knockout samples 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) were incubated overnight at 4°C at 1 in 1000 dilution and 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.
Allele-1: 5 bp deletion in exon 2
Allele-2: 2 bp deletion in exon 2
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab257066 has not yet been referenced specifically in any publications.