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AB258073

Human NFATC3 (NFAT4) knockout HEK-293T cell lysate

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NFATC3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

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

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Western blot - Human NFATC3 (NFAT4) knockout HEK-293T cell lysate (AB258073)
  • WB

Lab

Western blot - Human NFATC3 (NFAT4) knockout HEK-293T cell lysate (AB258073)

Lane 1 : Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate, 40 ug
Lane 2 : NFATC3 (NFAT4) knockout HEK293T (human embryonic kidney epithelial cell) whole cell lysate, 40 ug
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 (<a href='/en-us/products/cell-lines/human-nfatc3-nfat4-knockout-hek-293t-cell-line-ab267300'>ab267300</a>)

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

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Sanger Sequencing - Human NFATC3 (NFAT4) knockout HEK-293T cell lysate (AB258073)
  • Sanger seq

Unknown

Sanger Sequencing - Human NFATC3 (NFAT4) knockout HEK-293T cell lysate (AB258073)

Homozygous : 1 bp insertion in exon3

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

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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></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

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?

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Properties and storage information

Gene name
NFATC3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

The protein NFAT4 also known as NF-ATc3 or NFATc3 is a transcription factor with a molecular mass of approximately 140 kDa. It is part of the nuclear factor of activated T-cells (NFAT) family. This protein primarily localizes in the cytoplasm and nucleus and is especially abundant in immune cells like T-cells and various other cell types including neuronal tissues and cardiac cells.
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.

Abnormal NFAT4 activity links to immune-related conditions and cardiac hypertrophy. Its dysregulation associates with autoimmune diseases wherein it affects T-cell activation and inflammation. In cardiac disorders excessive activation of NFAT4 in conjunction with calcineurin contributes to maladaptive cardiac growth presenting a potential therapeutic target for hypertrophic heart conditions. The understanding of NFAT4's involvement in these diseases emphasizes its significance in developing targeted treatments.
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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

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB283690

Anti-NFAT4/NF-ATc3 antibody [EPR24994-68]

20ul selling size|RabMAb|Recombinant|KO Validated

Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-68] (AB283690)

  • 0
  • 0
Proteins & Peptides

AB114667

Recombinant Human NFAT4/NF-ATc3 protein

SDS-PAGE - Recombinant Human NFAT4/NF-ATc3 protein (AB114667)

  • 0
  • 0

Product promise

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