Human NKX2-5 knockout HEK-293 cell lysate
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- WB
Lab
Western blot - Human NKX2-5 knockout HEK-293 cell lysate (AB261716)
Lane 1 : Wild-type HEK-293 cell lysate 20 μg
Lane 2 : NKX2-5 knockout HEK-293 cell lysate 20 μg
Lane 3 : HeLa cell lysate 20 μg
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 °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-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 (<a href='/en-us/products/cell-lines/human-nkx2-5-knockout-hek-293-cell-line-ab261899'>ab261899</a>)
Lane 3:
HeLa cell lysate at 20 µg
Predicted band size: 35 kDa
Observed band size: 25 kDa,30 kDa,40 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human NKX2-5 knockout HEK-293 cell lysate (AB261716)
Knockout achieved by CRISPR/Cas9; X = 16 bp deletion, 5 bp deletion; Frameshift = 99.66%
Complementary Products
Reactivity data
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 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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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