Human PTPN2 knockout HCT116 cell line
- Advanced Validation
- What is this?
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PTPN2 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. 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
PTN2_HUMAN, PTPN 2, PTPT, Protein tyrosine phosphatase non receptor type 2, T-cell protein-tyrosine phosphatase, TCELLPTP, Tyrosine-protein phosphatase non-receptor type 2
- WB
Lab
Western blot - Human PTPN2 knockout HCT116 cell line (AB287729)
Western blot : Anti-PTPN2 antibody staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, anti-PTPN2 antibody was shown to bind specifically to PTPN2. A band was observed at 48 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in PTPN2 knockout cell line. To generate this image, wild-type and PTPN2 knockout HCT 116 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Anti-PTPN2 antibody at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human PTPN2 knockout HCT116 cell line (ab287729)
Lane 2:
PTPN2 knockout HCT 116 cell lysate at 20 µg
Lane 3:
Wild-type Jurkat cell lysate at 20 µg
Lane 4:
PTPN2 knockout Jurkat cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 48 kDa
false
- WB
Lab
Western blot - Human PTPN2 knockout HCT116 cell line (AB287729)
Western blot : Anti-PTPN2 antibody [EPR28199-34] (ab314496) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab314496 was shown to bind specifically to PTPN2. A band was observed at 48 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in PTPN2 knockout cell line. To generate this image, wild-type and PTPN2 knockout HCT 116 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-TCPTP antibody [EPR28199-34] (<a href='/en-us/products/primary-antibodies/tcptp-antibody-epr28199-34-ab314496'>ab314496</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human PTPN2 knockout HCT116 cell line (ab287729)
Lane 2:
PTPN2 knockout HCT 116 cell lysate at 20 µg
Lane 3:
Wild-type Jurkat cell lysate at 20 µg
Lane 4:
PTPN2 knockout Jurkat cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 48 kDa
false
- NGS
Lab
Next Generation Sequencing - Human PTPN2 knockout HCT116 cell line (AB287729)
91 bp deletion (allele 1), 2 bp insertion and 5 bp deletion (allele 2) in exon 6, CCDS11865.1
Reactivity data
Product details
Recommended control: Human wild-type HCT116 cell line (ab288559). 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.
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
McCoY5a + 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
TCPTP plays a role in the control of cell growth differentiation and immune response. It functions independently not as part of a larger complex. This enzyme acts as a negative regulator of signaling pathways by dephosphorylating proteins that drive these cellular processes. Its activity ensures a balance in cellular signaling preventing overactivation which could lead to uncontrolled cell proliferation or inappropriate immune responses.
Pathways
TCPTP holds significant function in the JAK-STAT and insulin signaling pathways. Within the JAK-STAT pathway TCPTP dephosphorylates members such as JAK1 and JAK3 helping modulate cytokine responses and immune regulation. In the insulin signaling pathway it influences the dephosphorylation of the insulin receptor thereby impacting insulin sensitivity. Such interactions demonstrate its involvement in key cellular processes necessary for immune function and metabolic regulation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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