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AB261899

Human NKX2-5 knockout HEK-293 cell line

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NKX2-5 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 16 bp deletion, 5 bp deletion; Frameshift = 99.66%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 Images
Western blot - Human NKX2-5 knockout HEK-293 cell line (AB261899)
  • WB

Lab

Western blot - Human NKX2-5 knockout HEK-293 cell line (AB261899)

False colour image of Western blot : Anti-Nkx2.5 antibody [EPR20168] 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 ab205263 was shown to bind specifically to Nkx2.5. A band was observed at 40 30 25 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in NKX2-5 knockout cell line ab261899 (knockout cell lysate ab261716). Bands at 37 and 35 kDa are non-specific believed to be NKX2-2 and NKX2-6. To generate this image wild-type and NKX2-5 knockout HEK-293 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 °. 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-Nkx2.5 antibody [EPR20168] (<a href='/en-us/products/primary-antibodies/nkx25-antibody-epr20168-ab205263'>ab205263</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

NKX2-5 knockout HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human NKX2-5 knockout HEK-293 cell line (ab261899)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 25 kDa,30 kDa,40 kDa

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Next Generation Sequencing - Human NKX2-5 knockout HEK-293 cell line (AB261899)
  • NGS

Supplier Data

Next Generation Sequencing - Human NKX2-5 knockout HEK-293 cell line (AB261899)

Knockout achieved by CRISPR/Cas9; X = 16 bp deletion, 5 bp deletion; Frameshift = 99.66%

Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 16 bp deletion, 5 bp deletion; Frameshift = 99.66%

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Complementary Products

Primary Antibodies

AB205263

Anti-Nkx2.5 antibody [EPR20168]

RabMAb|Recombinant|KO Validated

Western blot - Anti-Nkx2.5 antibody [EPR20168] (AB205263)

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Cell Lines & Lysates

AB261716

Human NKX2-5 knockout HEK-293 cell lysate

Western blot - Human NKX2-5 knockout HEK-293 cell lysate (AB261716)

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Primary Antibodies

AB92574

Anti-CD90 / Thy1 antibody [EPR3132]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD90 / Thy1 antibody [EPR3132] (AB92574)

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Proteins & Peptides

AB152305

Recombinant Human Nkx2.5 protein

SDS-PAGE - Recombinant Human Nkx2.5 protein (AB152305)

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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

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
NKX2-5
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

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

Nkx2.5 also known as NKX2-5 or CSX is a homeobox-containing transcription factor with a molecular mass of approximately 35 kDa. It is primarily expressed in the heart specifically in cardiac progenitor cells and also found in low levels in the spleen stomach and pancreas. As a transcription factor Nkx2.5 plays a significant role in regulating the expression of genes critical for cardiac development and differentiation. The protein's expression is tightly regulated during early embryogenesis and maintained in the heart throughout life.
Biological function summary

Nkx2.5 functions to direct the development of cardiac myocytes by regulating genes involved in cell specification and tissue formation. Nkx2.5 does not solely operate but interacts with other cardiac-specific transcription factors such as GATA4 and TBX5 collectively forming a transcriptional complex. This cooperative interaction is essential in orchestrating the genetic program necessary for heart morphogenesis and cellular proliferation within the developing myocardium.

Pathways

The protein Nkx2.5 significantly influences cardiogenesis and heart formation pathways. Its activity is necessary in the Wnt signaling pathway important for the initial stages of cardiac cell lineage differentiation. Additionally Nkx2.5 works in tandem with the protein MEF2C to regulate cardiac-specific genes ensuring the proper development and functioning of cardiac tissues. Both pathways highlight important regulatory networks where Nkx2.5 is an important player in heart development.

Mutations or dysregulations of Nkx2.5 can lead to congenital heart defects such as atrial septal defects and tetralogy of Fallot. These conditions often arise due to disrupted genetic signaling during heart formation. Nkx2.5 mutations have been associated with conduction system defects in conjunction with the GATA4 protein both known to be involved in maintaining normal cardiac structure and function. Understanding Nkx2.5's role in these diseases aides in developing potential therapeutic approaches targeting congenital heart abnormalities.
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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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Target data

See full target information NKX2-5
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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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