Human NFATC2 knockout Raji cell line
- Advanced Validation
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NFATC2 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 122 bp deletion and 3 bp insertion in exon 2.
View Alternative Names
AI607462, KIAA0611, NF-ATc2, NF-ATp, NFAC2_HUMAN, NFAT pre-existing subunit, NFAT transcription complex, preexisting component, NFAT1-D, Nuclear factor of activated T cells cytoplasmic 2, Nuclear factor of activated T cells cytoplasmic calcineurin dependent 2, Nuclear factor of activated T cells pre-existing component, Nuclear factor of activated T-cells, Preexisting nuclear factor of activated T cells 2, T-cell transcription factor NFAT1, cytoplasmic 2
- WB
Lab
Western blot - Human NFATC2 knockout Raji cell line (AB280906)
False colour image of Western blot : Anti-NFAT1 antibody [EPR24658-149] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283649 was shown to bind specifically to NFAT1. A band was observed at 100 kDa in wild-type Raji cell lysates with no signal observed at this size in NFATC2 knockout cell line ab280906 (knockout cell lysate ab282940). The band observed in the knockout lysate lane below 100 kDa is likely to represent a truncated form of NFAT1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NFATC2 knockout Raji 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Lanes 1 and 3:
Western blot - Anti-NFAT1 antibody [EPR24658-149] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/nfat1-antibody-epr24658-149-bsa-and-azide-free-ab283659'>ab283659</a>) at 1/1000 dilution
Lanes 2 and 4:
Western blot - Anti-NFAT1 antibody [EPR24658-149] (<a href='/en-us/products/primary-antibodies/nfat1-antibody-epr24658-149-ab283649'>ab283649</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji cell lysate at 20 µg
Lane 2:
NFATC2 CRISPR-Cas9 edited Raji cell lysate at 20 µg
Lane 2:
Western blot - Human NFATC2 knockout Raji cell line (ab280906)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 100 kDa
Observed band size: 100 kDa
false
- WB
Lab
Western blot - Human NFATC2 knockout Raji cell line (AB280906)
False colour image of Western blot : Anti-NFAT1 antibody [EPR24658-43] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283691 was shown to bind specifically to NFAT1. A band was observed at 100 kDa in wild-type Raji cell lysates with no signal observed at this size in NFATC2 knockout cell line ab280906 (knockout cell lysate ab282940). The band observed in the knockout lysate lane below 100 kDa is likely to represent a truncated form of NFAT1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NFATC2 knockout Raji 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-NFAT1 antibody [EPR24658-43] (<a href='/en-us/products/primary-antibodies/nfat1-antibody-epr24658-43-ab283691'>ab283691</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji cell lysate at 20 µg
Lane 2:
NFATC2 CRISPR-Cas9 edited Raji cell lysate at 20 µg
Lane 2:
Western blot - Human NFATC2 knockout Raji cell line (ab280906)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 100 kDa
Observed band size: 100 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human NFATC2 knockout Raji cell line (AB280906)
122 bp deletion and 3 bp insertion in exon 2
Reactivity data
Product details
Recommended Control: Human wild-type Raji (ab275473). 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.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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 for bath 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 - 5x105 cells/mL(for initial passages it is recomended to culture the cells in the higher range of recomended seeding density). 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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 4x105 cells/mL is recommended.
- A maximum of 3x106 viable cells/mL is obtainable.
Culture medium
RPMI + 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
NFAT1 influences immune response and development. It acts as part of a large complex with other transcription factors to bind DNA and regulate transcription of cytokines and other critical immune molecules. In activated T-cells NFAT1 controls the expression of genes important for immune function including interleukins. In neuronal tissues it contributes to neural growth and survival highlighting its multifunctional nature in different cell types.
Pathways
NFAT1 integrates into the calcineurin signaling pathway and plays a significant role in T-cell activation. It acts downstream of the calcium signaling cascade which leads to dephosphorylation and activation by the phosphatase calcineurin. NFAT1 works closely with proteins like AP-1 and GATA to facilitate the transcription of immune-related genes. These interactions highlight NFAT1's importance in immune signaling pathways which modulate immune surveillance and homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Suspension
Gender
Male
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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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