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AB282940

Human NFATC2 knockout Raji cell lysate

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NFATC2 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 122 bp deletion and 3 bp insertion in exon 2.
3 Images
Western blot - Human NFATC2 knockout Raji cell lysate (AB282940)
  • WB

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Western blot - Human NFATC2 knockout Raji cell lysate (AB282940)

Lane 1 : Wild-type Raji cell lysate 20 μg
Lane 2 : NFATC2 knockout Raji cell lysate 20 μg
Lane 3 : Jurkat cell lysate 20 μg
Lane 4 : SH-SY5Y cell lysate 20 μg
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.

All lanes:

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 knockout Raji cell lysate at 20 µg

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

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Western blot - Human NFATC2 knockout Raji cell lysate (AB282940)
  • WB

Lab

Western blot - Human NFATC2 knockout Raji cell lysate (AB282940)

Lane 1 : Wild-type Raji cell lysate 20 μg
Lane 2 : NFATC2 knockout Raji cell lysate 20 μg
Lane 3 : Jurkat cell lysate 20 μg
Lane 4 : SH-SY5Y cell lysate 20 μg
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 knockout Raji cell lysate at 20 µg

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

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Sanger Sequencing - Human NFATC2 knockout Raji cell lysate (AB282940)
  • Sanger seq

Lab

Sanger Sequencing - Human NFATC2 knockout Raji cell lysate (AB282940)

122 bp deletion and 3 bp insertion in exon 2

Key facts

Cell type

Raji

Species or organism

Human

Tissue

Lymphatic

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 122 bp deletion and 3 bp insertion in exon 2

Disease

Lymphoma

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Complementary Products

Primary Antibodies

AB2722

Anti-NFAT1 antibody [25A10.D6.D2]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT1 antibody [25A10.D6.D2] (AB2722)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }
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Product details

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab282940-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type Raji cell lysate", "number":"AB282940-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human NFATC2 knockout Raji cell lysate", "number":"AB282940-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
NFATC2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

NFAT1 also known as NFATc2 is a transcription factor involved in gene regulation with a molecular weight of approximately 100 kDa. It belongs to the NFAT (nuclear factor of activated T-cells) family and is expressed in a variety of tissues including the immune system and the nervous system. NFAT1 plays an essential role in activating specific gene expressions by translocating into the nucleus upon cell stimulation. Researchers frequently use NFAT molecular weight to confirm its presence in protein analyses such as NFAT western blot techniques.
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.

NFAT1 associates with autoimmune diseases and certain cancers. In autoimmune conditions such as rheumatoid arthritis aberrant NFAT1 function leads to excessive cytokine production exacerbating inflammation. The protein also links to cancer development particularly in lymphoid malignancies where dysregulated NFAT1 activity contributes to uncontrolled cell proliferation. It interacts with oncogenic kinases and other dysregulated proteins in these disorders influencing both their progression and possible therapeutic targeting.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Suspension

Gender

Male

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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