Human NFATC3 (NFAT4) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(1 Publication)
NFATC3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3. 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
C80703, D8Ertd281e, NF-AT4, NF-ATc3, NFAC3_HUMAN, NFATx, Nuclear factor of activated T cells, cytoplasmic 3, Nuclear factor of activated T-cells, T-cell transcription factor NFAT4, cytoplasmic 3, nuclear factor of activated T cells, cytoplasmic, calcineurin dependent 3
- WB
Lab
Western blot - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)
Lanes 1- 2 : Merged signal (red and green).Green - ab283690 observed at 140 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab283690 was shown to react with NFATC3 (NFAT4) in wild-type HEK293T cells in western blot. Loss of signal was observed when NFATC3 (NFAT4) knockout cell line ab267300 (knockout cell lysate ab258073) was used. Wild-type HEK293T and NFATC3 (NFAT4) knockout HEK293T 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. ab283690 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1/1000 dilution and 1/20,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Lysates were made freshly and used in WB immediately to minimize protein degradation.
Blocking and diluting buffer and concentration : 3% NFDM/TBST
All lanes:
Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-68] (<a href='/en-us/products/primary-antibodies/nfat4-nf-atc3-antibody-epr24994-68-ab283690'>ab283690</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 40 µg
Lane 2:
NFATC3 (NFAT4) knockout HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 40 µg
Lane 2:
Western blot - Human NFATC3 (NFAT4) knockout HEK-293T cell line (ab267300)
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 116 kDa
Observed band size: 140 kDa
false
- Cell Culture
Unknown
Cell Culture - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)
Representative images of NFATC3 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 XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)
Homozygous : 1 bp insertion in exon3
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
Zygosity
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
NFAT4/NF-ATc3 acts as a regulator of gene expression by binding to DNA in the nucleus. It forms part of the NFAT complex working with other members of this family to regulate genes involved in cell differentiation and development. In immune cells this protein plays a role in mediating responses to external signals influencing cytokine production and aiding the regulation of immune responses. Other tissues including neurons and cardiac cells also rely on NFAT4 for specific signaling functions related to cell growth and survival.
Pathways
NFAT4/NF-ATc3 is involved in key signaling mechanisms notably the calcium signaling pathway. This pathway works closely with other proteins such as calcineurin which dephosphorylates NFAT proteins leading to their activation and nuclear translocation. Additionally interactions within the Ras/MAPK pathway highlight NFAT4's role in cell proliferation and differentiation processes interconnecting with various signaling proteins that drive these pathways.
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
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Cell biology international 47:480-491 PubMed36273427
2022
Applications
Unspecified application
Species
Unspecified reactive species
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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