CHI3L1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 7.
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Key facts
- Cell type
THP-1
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 7
Alternative names
39 kDa synovial protein, ASRT7, CGP-39, CH3L1_HUMAN, CHI3L1, Cartilage glycoprotein 39, Chitinase 3 like protein 1 precursor, Chitinase-3-like protein 1, Chondrocyte protein YKL40, GP-39, HCGP 3P, YKL-40, YYL 40, chitinase, chitinase 3 like 1, chitinase 3 like 1 (cartilage glycoprotein 39), hCGP-39
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CHI3L1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 7.
Key facts
- Cell type
THP-1
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 7
- Disease
Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
CHI3L1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
- Zygosity
Homozygous
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
Recommended control: Human wild-type THP-1 cell line (Human wild-type THP-1 cell line ab281894). 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
YKL-40 also known as chitinase-3-like protein 1 (CHI3L1) is a glycoprotein with a molecular weight of approximately 40 kDa. Scientists often study it for its roles in inflammatory processes. It has expressions in numerous tissues including liver cartilage and the central nervous system. The CHI3L1 protein does not possess chitinase activity unlike other members of the chitinase family yet it interacts with various ligands to influence cellular behavior.
- Biological function summary
YKL-40 plays significant roles in tissue remodeling and inflammation. It does not form part of a larger complex by itself but influences several cellular processes. This protein affects cell proliferation migration and survival making it involved in both normal physiological processes and pathological conditions. CHI3L1 also exhibits a connection to the immune response where it modulates macrophage activity and interacts with other immune cells.
- Pathways
YKL-40 is integral to the inflammatory and extracellular matrix pathways. It associates with proteins such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) influencing signaling cascades that modulate inflammation. Through these interactions CHI3L1 becomes a mediator of connective tissue metabolism and immune reactions in various pathophysiological states.
- Associated diseases and disorders
YKL-40 shows strong links to conditions like osteoarthritis and certain cancers. In osteoarthritis elevated levels of CHI3L1 reflect increased cartilage degradation while in oncology it correlates with tumor angiogenesis and metastasis potential. Its connection with proteins such as matrix metalloproteinases highlights its role in disease mechanisms highlighting its potential as a biomarker for early detection and disease progression monitoring.
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3 product images
Western blot - Human CHI3L1 knockout THP-1 cell line (ab280038)
False colour image of Western blot: Anti-YKL-40/CHI3L1 antibody [EPR19078-157] 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-YKL-40/CHI3L1 antibody [EPR19078-157] ab255297 was shown to bind specifically to YKL-40/CHI3L1. A band was observed at 37/40 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CHI3L1 knockout cell line ab280038 (knockout cell lysate Human CHI3L1 knockout THP-1 cell lysate ab280097). To generate this image, wild-type and CHI3L1 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 °C. 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.
All lanes: Western blot - Anti-YKL-40/CHI3L1 antibody [EPR19078-157] (Anti-YKL-40/CHI3L1 antibody [EPR19078-157] ab255297) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: CHI3L1 knockout THP-1 cell lysate at 20 µg
Lane 3: U-87 MG cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 37 kDa, 40 kDa
Western blot - Human CHI3L1 knockout THP-1 cell line (ab280038)
False colour image of Western blot: Anti-YKL-40/CHI3L1 antibody staining at 1 ug/ml, 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-YKL-40/CHI3L1 antibody ab77528 was shown to bind specifically to YKL-40/CHI3L1. A band was observed at 33-42 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CHI3L1 knockout cell line ab280038 (knockout cell lysate Human CHI3L1 knockout THP-1 cell lysate ab280097). To generate this image, wild-type and CHI3L1 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 °C. 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.
All lanes: Western blot - Anti-YKL-40/CHI3L1 antibody (Anti-YKL-40/CHI3L1 antibody ab77528) at 1 µg/mL
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: CHI3L1 knockout THP-1 cell lysate at 20 µg
Lane 3: U-87 MG cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 33-42 kDa
Sanger Sequencing - Human CHI3L1 knockout THP-1 cell line (ab280038)
14 bp deletion in exon 7
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