Human PTPN11 (SHP2) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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PTPN11 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
BPTP3, JMML, METCDS, MGC14433, OTTHUMP00000166107, OTTHUMP00000166108, PTN11_HUMAN, PTP-1D, PTP-2C, PTPN11, Protein tyrosine phosphatase 2, Protein tyrosine phosphatase non receptor type 11, Protein-tyrosine phosphatase 1D, Protein-tyrosine phosphatase 2C, SAP-2, SH-PTP2, SH-PTP3, SH2 domain containing protein tyrosine phosphatase 2, SHP-2, Tyrosine-protein phosphatase non-receptor type 11
- WB
Lab
Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
Lanes 1- 2 : Merged signal (red and green). Green - ab187040 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab187040 was shown to react with SHP2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266450 (knockout cell lysate ab257618) was used. Wild-type HEK-293T and PTPN11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187040 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 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.
All lanes:
Western blot - Anti-SHP2 antibody [EPR17829-9] (<a href='/en-us/products/primary-antibodies/shp2-antibody-epr17829-9-ab187040'>ab187040</a>) at 1/5000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PTPN11 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (ab266450)
Predicted band size: 68 kDa
Observed band size: 68 kDa
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- WB
Lab
Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
Lanes 1- 2 : Merged signal (red and green). Green - ab32083 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32083 was shown to react with SHP2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266450 (knockout cell lysate ab257618) was used. Wild-type HEK-293T and PTPN11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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.
All lanes:
Western blot - Anti-SHP2 antibody [Y478] (<a href='/en-us/products/primary-antibodies/shp2-antibody-y478-ab32083'>ab32083</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PTPN11 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (ab266450)
Predicted band size: 68 kDa
Observed band size: 68 kDa
false
- Cell Culture
Lab
Cell Culture - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
Representative images PTPN11 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
Allele-1 : 2 bp deletion in exon1
- Sanger seq
Unknown
Sanger Sequencing - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
Allele-2 : 1 bp insertion in exon 1.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human PTPN11 (SHP2) knockout HEK-293T cell line (AB266450)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PTPN11 KO HEK293T (PTPN11 KO knock out human embryonic kidney epithelial cell) (ab266450) cells labelling SHP2 with ab300579 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in wildtype HEK293T cell line, while showing no staining in PTPN11 knockout HEK293T cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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 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
DMEM (High Glucose) + 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
The role of SHP2 extends to involvement in several signaling cascades such as the Ras/MAPK and PI3K/AKT pathways. It functions as an essential component within protein complexes that facilitate cell communication and response to external signals. The protein modulates growth factor signaling and cytokine signaling highlighting its significance in normal cell function and development. SHP2's statement in signaling processes makes it an important regulator of cellular dynamics.
Pathways
SHP2 participates in the Ras/MAPK and PI3K/AKT signaling pathways which are important for regulating cell growth survival and differentiation. Within these pathways SHP2 interacts with various signaling molecules including Grb2 Sos and Gab family adaptors. These interactions coordinate cellular responses to growth factors and other extracellular cues ensuring proper pathway activation and control. By serving as a critical mediator SHP2 integrates signals that are necessary for appropriate cellular outcomes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB187040
Anti-SHP2 antibody [EPR17829-9]
RabMAb|Recombinant|KO Validated|Trial Size
![Immunoprecipitation - Anti-SHP2 antibody [EPR17829-9] (AB187040)](https://content.abcam.com/products/images/shp2-antibody-epr17829-9-ab187040--immunoprecipitation-img298.jpg)
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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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