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AB257805

Human YWHAG (14-3-3 gamma) knockout HEK-293T cell lysate

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YWHAG 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 exon1.

View Alternative Names

14 3 3 gamma, 14 3 3 protein gamma subtype, 14-3-3 protein gamma, 1433G_HUMAN, 3 monooxygenase/tryptophan 5 monooxgenase activation protein gamma polypeptide, N-terminally processed, Protein kinase C inhibitor protein 1, Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein gamma polypeptide, Ywhag

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Western blot - Human YWHAG (14-3-3 gamma) knockout HEK-293T cell lysate (AB257805)
  • WB

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Western blot - Human YWHAG (14-3-3 gamma) knockout HEK-293T cell lysate (AB257805)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : YWHAG knockout HEK293T cell lysate (20 ug)
Lane 3 : A431 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab137048 was shown to specifically react with 14-3-3 gamma/YWHAG in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266734 (knockout cell lysate ab257805) was used. Wild-type and 14-3-3 gamma/YWHAG knockout samples were subjected to SDS-PAGE. ab137048 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 dilution and 1 in 20000 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-14-3-3 gamma/YWHAG antibody [EPR9654(B)] (<a href='/en-us/products/primary-antibodies/14-3-3-gamma-ywhag-antibody-epr9654b-ab137048'>ab137048</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

YWHAG knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human YWHAG (14-3-3 gamma) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ywhag-14-3-3-gamma-knockout-hek-293t-cell-line-ab266734'>ab266734</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 28 kDa

Observed band size: 28 kDa

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Sanger Sequencing - Human YWHAG (14-3-3 gamma) knockout HEK-293T cell lysate (AB257805)
  • Sanger seq

Unknown

Sanger Sequencing - Human YWHAG (14-3-3 gamma) knockout HEK-293T cell lysate (AB257805)

Homozygous : 1 bp insertion in exon1

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 exon1.

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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
YWHAG
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.

14-3-3 gamma also known as YWHAG or 14-3-3 gamma protein is a member of the 14-3-3 protein family. These proteins play significant roles in signal transduction. 14-3-3 gamma is a dimeric protein with a molecular mass of about 28 to 32 kDa per monomer. It is ubiquitously expressed including in tissues such as brain muscle and heart. This protein influences a wide range of cellular processes by interacting with various signaling proteins often through recognition of phosphorylated serine or threonine motifs.
Biological function summary

Proteins within the 14-3-3 family like the 14-3-3 gamma function as adaptors and scaffolds in signal transduction pathways. They are part of multi-protein complexes that modulate interactions among proteins within the cell. 14-3-3 gamma interacts dynamically with a variety of target proteins assisting in their activities and proper localization. Its interaction facilitates cellular processes including cell cycle control apoptosis and stress response owing to its activity as a regulator of distinct signaling pathways.

Pathways

14-3-3 gamma participates in the MAPK/ERK pathway which is pivotal for cell growth and differentiation. It also plays an integral role in the PI3K/AKT pathway which contributes to survival and proliferation signals. In these pathways 14-3-3 gamma interacts with important kinases such as Raf and AKT. These interactions help transduce signals from receptors on the cell surface to the nucleus impacting gene expression and cellular responses.

Research has linked 14-3-3 gamma to neurodegenerative diseases such as Alzheimer's disease. In Alzheimer's 14-3-3 gamma interacts with proteins like tau which become abnormally phosphorylated and aggregate. Additionally this protein has associations with certain cancers where it might influence tumor growth and progression through interactions with cell cycle regulatory proteins. Its modulation of key signaling pathways makes it a candidate of interest in the development of therapeutic strategies for these diseases.
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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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