CTSS KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%.
Images
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%
Alternative names
CATS_HUMAN, CTSS, Cathepsin S
Recommended products
CTSS KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%.
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%
- Disease
- Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
- CTSS
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Suspension
- 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
- Culture medium
- RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
- 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 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
Cathepsin S sometimes referred to as CTSS is a cysteine protease enzyme weighing about 24-29 kDa part of the papain family. It occurs mainly in lysosomes and is expressed in a variety of cells including antigen-presenting cells like macrophages dendritic cells and B lymphocytes. Cathepsin S functions to cleave proteins at lysines playing a central role in protein degradation and antigen processing. It distinguishes itself from other cathepsins by maintaining activity in an alkaline environment and is essential for the processing of invariant chain (Ii) in MHC class II molecules.
- Biological function summary
Cathepsin S participates in immune system modulation and matrix degradation. It stands out in its role in the immune response cleaving antigenic proteins for presentation by MHC class II therefore influencing T-cell activation. Although synonymous with lysosomal activity it also operates extracellularly contributing to tissue remodeling and inflammation. Cathepsin S is not part of a larger complex but interacts with MHC class II impacting antigen presentation.
- Pathways
This protease is critical in the immune system and extracellular matrix degradation pathways. In the context of the immune pathway CTSS interacts with other proteases such as Cathepsin L and Cathepsin B to perform antigen trimming and processing. Moreover CTSS supports the activation of T lymphocytes by ensuring the correct presentation of peptide fragments on MHC class II molecules. In matrix degradation CTSS indirectly influences collagen breakdown and interaction with other matrix metalloproteinases.
- Associated diseases and disorders
Cathepsin S is associated with autoimmune diseases like rheumatoid arthritis where it contributes to tissue inflammation and joint damage. It also relates to atherosclerosis promoting instability in atherosclerotic plaques which could result in heart disease. In rheumatoid arthritis interactions with cytokines like TNF-alpha amplify inflammatory responses while in atherosclerosis CTSS links with other proteases to degrade extracellular matrix components leading to vulnerable lesions.
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2 product images
Western blot - Human CTSS knockout THP-1 cell line (ab275846)
False colour image of Western blot: Anti-Cathepsin S antibody [EPR5128] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-Cathepsin S antibody [EPR5128] ab134157 was shown to bind specifically to Cathepsin S. A band was observed at 25 kDa in wild-type THP-1 cell lysates with no signal observed at this size in Ctss knockout cell line ab275846 (knockout cell lysate ab275820). To generate this image wild-type and Ctss knockout THP-1 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°. 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.
Lanes 1 - 2: Western blot - Anti-Cathepsin S antibody [EPR5128] (Anti-Cathepsin S antibody [EPR5128] ab134157) at 1/1000 dilution
Lanes 3 - 4: Western blot - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free ab248706) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: Ctss knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human CTSS knockout THP-1 cell line (ab275846)
Lane 3: U-87 MG cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 25 kDa
Next Generation Sequencing - Human CTSS knockout THP-1 cell line (ab275846)
Knockout achieved by CRISPR/Cas9; X = 14 bp deletion; Frameshift: 100%
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