Human CTSS knockout THP-1 cell line
- Advanced Validation
- What is this?
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CTSS KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
CATS_HUMAN, CTSS, Cathepsin S
- WB
Lab
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] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Lanes 1 - 2:
Western blot - Anti-Cathepsin S antibody [EPR5128] (<a href='/en-us/products/primary-antibodies/cathepsin-s-antibody-epr5128-ab134157'>ab134157</a>) at 1/1000 dilution
Lanes 3 - 4:
Western blot - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/cathepsin-s-antibody-epr5128-bsa-and-azide-free-ab248706'>ab248706</a>) 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
Predicted band size: 37 kDa
Observed band size: 25 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human CTSS knockout THP-1 cell line (AB275846)
Knockout achieved by CRISPR/Cas9; X = 14 bp deletion; Frameshift : 100%
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. 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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB134157
Anti-Cathepsin S antibody [EPR5128]
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin S antibody [EPR5128] (AB134157)](https://content.abcam.com/products/images/cathepsin-s-antibody-epr5128-ab134157--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img177660.jpg)
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Cell Lines & Lysates
AB276116
Human CASP1 (Caspase-1) knockout THP-1 cell line
Advanced Validation
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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