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AB267300

人NFATC3 (NFAT4) knockout HEK-293T cell line

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

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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 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

查看别名

NFATc3, NFATx, NFAT4, NFATC3, NF-AT4c, T-cell transcription factor NFAT4, NF-ATc3, NF-AT4

3 Images
Western blot - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)
  • 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='/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='/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='/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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Cell Culture - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)
  • 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 Sequencing - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)
  • Sanger seq

Unknown

Sanger Sequencing - Human NFATC3 (NFAT4) knockout HEK-293T cell line (AB267300)

Homozygous : 1 bp insertion in exon3

关键信息

细胞类型

HEK-293T

种属

Human

组织

Kidney

形式

Liquid

form

敲除验证

Sanger Sequencing,Western blot

突变描述

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3

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搭配使用产品

Cell Lines & Lysates

AB266742

Human GSK3A (GSK3 alpha) knockout HEK-293T cell line

Advanced Validation

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

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Primary Antibodies

AB99431

Anti-NFATC4 antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFATC4 antibody (AB99431)

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Cell Lines & Lysates

AB265070

Human FBXW7 knockout HEK-293T cell line

Sanger Sequencing - Human FBXW7 knockout HEK-293T cell line (AB265070)

  • 0
  • 0
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)

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  • 0
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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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产品详情

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.

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规格

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性能和储存信息

基因名称
NFATC3
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Sanger Sequencing, Western blot
合子性
Homozygous
运输条件
Dry Ice
推荐的短期储存条件
-196°C
推荐的长期储存条件
-196°C
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补充信息

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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质量控制

STR 分析

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

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细胞培养

生物安全等级

EU: 2 US: 2

贴壁/悬浮

Adherent

性别

Female

初始处理指南

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

传代培养指南
  • 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.
培养基

DMEM (High Glucose) + 10% FBS

低温储藏试剂

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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产品实验方案

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靶点信息

See full target information NFATC3
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文献 (1)

Recent publications for all applications. Explore the full list and refine your search

Cell biology international 47:480-491 PubMed36273427

2022

Inhibition of intermittent calcium-activated potassium channel (SK4) attenuates Ang II-induced hypertrophy of human-induced stem cell-derived cardiomyocytes via targeting Ras-Raf-MEK1/2-ERK1/2 and CN-NFAT signaling pathways.

Applications

Unspecified application

Species

Unspecified reactive species

Hongyi Zhao,Xi Wang,Yanhong Tang,Qingyan Zhao,Congxin Huang
View all publications
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