Human TNC (Tenascin C) knockout U-2 OS cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human TNC (Tenascin C) knockout U-2 OS cell line (AB262488)
False colour image of Western blot : Anti-Tenascin C antibody [EB2] staining at 1/1000 dilution shown in green; Rabbit anti-ACTN4 [EPR2533(2)] (ab108198) loading control staining at 1/20000 dilution shown in red. In Western blot ab88280 was shown to bind specifically to Tenascin C. A band was observed at 200/270 kDa in treated wild-type U-2 OS cell lysates with no signal observed at this size in TNC knockout cell line ab262488 (knockout cell lysate ab263924). To generate this image wild-type and TNC knockout U-2 OS cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 2 % BSA 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 before development with Optiblot (ECL reagent ab133456) and imaged with 16 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Mouse (H+L) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-Tenascin C antibody [EB2] (<a href='/en-us/products/primary-antibodies/tenascin-c-antibody-eb2-ab88280'>ab88280</a>) at 1/1000 dilution
Lane 1:
Wild-type U-2 OS Treated BFA (5 ug/mL, 6 h) cell lysate at 40 µg
Lane 2:
TNC knockout U-2 OS Treated BFA (5 ug/mL, 6 h) cell lysate at 40 µg
Lane 2:
Western blot - Human TNC (Tenascin C) knockout U-2 OS cell line (ab262488)
Lane 3:
Wild-type U-2 OS Vehicle Control BFA (0 ug/mL, 6 h) cell lysate at 40 µg
Lane 4:
TNC knockout U-2 OS Vehicle Control BFA (0 ug/mL, 6 h) cell lysate at 40 µg
Lane 5:
U-87 MG Treated BFA (5ug/mL, 6h) cell lysate at 40 µg
Lane 6:
U-87 MG cell lysate at 40 µg
Predicted band size: 241 kDa
Observed band size: 200 kDa,270 kDa
false
- WB
Lab
Western blot - Human TNC (Tenascin C) knockout U-2 OS cell line (AB262488)
Western blot : Anti-TNC antibody [EPR4219] (ab108930) staining at 1/500 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108930 was shown to bind specifically to TNC. A band was observed at 122 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in TNC knockout cell line. To generate this image, wild-type and TNC knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 2 % BSA 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 before development with a high-sensitivity ECL substrate kit and imaged with 5 seconds exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Tenascin C antibody [EPR4219] (<a href='/en-us/products/primary-antibodies/tenascin-c-antibody-epr4219-ab108930'>ab108930</a>) at 1/500 dilution
Lane 1:
Wild-type U-2 OS BFA (5 ug/mL, 6 h) cell lysate at 40 µg
Lane 2:
Wild-type U-2 OS BFA (0 ug/mL, 6 h) cell lysate at 40 µg
Lane 3:
TNC knockout U-2 OS BFA (5 ug/mL, 6 h) cell lysate at 40 µg
Lane 4:
TNC knockout U-2 OS BFA (0 ug/mL, 6 h) cell lysate at 40 µg
Lane 5:
U-87 MG Treated BFA (5 ug/mL, 6 h) cell lysate at 10 µg
Lane 6:
U-87 MG Control BFA (0 ug/mL, 6 h) cell lysate at 10 µg
Secondary
All lanes:
HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human TNC (Tenascin C) knockout U-2 OS cell line (AB262488)
29 bp deletion after Asp201 of the WT protein
Complementary Products
Primary Antibodies
AB108930
Anti-Tenascin C antibody [EPR4219]
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Frozen sections) - Anti-Tenascin C antibody [EPR4219] (AB108930)](https://content.abcam.com/products/images/tenascin-c-antibody-epr4219-ab108930--immunohistochemistry-frozen-sections-img110582.jpg)
- 4
- 16
Cell Lines & Lysates
AB266883
Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line
Advanced Validation
- 0
- 0
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 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
The large hexamer known as Tenascin C plays a significant role in modulating cell adhesion migration and proliferation especially during developmental processes and injury responses. It incorporates into the existing extracellular matrix as a loose network which provides elasticity and supports cellular activities. This feature allows Tenascin C to interact with other matrix molecules and cell surface receptors to regulate tissue morphogenesis.
Pathways
Tenascin C actively participates in the Wnt and TGF-beta signaling pathways important for controlling cellular behavior in development and regeneration. It associates with integrins and glycoproteins to influence cell responses to signaling events in these pathways. By modulating these interactions Tenascin C can impact processes such as epithelial-to-mesenchymal transition and angiogenesis affecting tissue architecture and function.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com