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AB256835

Human AKT1 knockout HeLa cell lysate

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AKT1 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 exon 2 and Insertion of the selection cassette in exon 2.
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Sanger Sequencing - Human AKT1 knockout HeLa cell lysate (AB256835)
  • Sanger seq

Unknown

Sanger Sequencing - Human AKT1 knockout HeLa cell lysate (AB256835)

Allele-2 : Insertion of the selection cassette in exon 2

Sanger Sequencing - Human AKT1 knockout HeLa cell lysate (AB256835)
  • Sanger seq

Unknown

Sanger Sequencing - Human AKT1 knockout HeLa cell lysate (AB256835)

Allele-1 : 1 bp insertion in exon 2

Sanger Sequencing - Human AKT1 knockout HeLa cell lysate (AB256835)
  • Sanger seq

Unknown

Sanger Sequencing - Human AKT1 knockout HeLa cell lysate (AB256835)

Allele-3 : Insertion of the selection cassette in exon 2

Western blot - Human AKT1 knockout HeLa cell lysate (AB256835)
  • WB

Lab

Western blot - Human AKT1 knockout HeLa cell lysate (AB256835)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : AKT1 knockout HeLa cell lysate (20µg)

Lane 3 : MCF7 cell lysate (20µg)

Lane 4 : Daudi cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab108202 observed at 60 kDa. Red - loading control ab181602 observed at 37 kDa.

ab108202 Anti-AKT1 antibody [M1843] was shown to specifically react with AKT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264852 (knockout cell lysate ab256835) was used. Wild-type and AKT1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108202 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-AKT1 antibody [M1843] (<a href='/en-us/products/primary-antibodies/akt1-antibody-m1843-ab108202'>ab108202</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human AKT1 knockout HeLa cell lysate (ab256835) at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 55 kDa

Observed band size: 60 kDa

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Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.

Disease

Adenocarcinoma

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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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256835-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB256835-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human AKT1 knockout HeLa cell lysate", "number":"AB256835-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
AKT1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Quality control - STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
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.

AKT1 also known as Protein Kinase B (PKB) is a serine/threonine kinase involved in various cellular processes. AKT1 plays an important role in mediating signals for cell survival growth and metabolism. This protein has a molecular weight of approximately 56 kDa. AKT1 is ubiquitously expressed in many tissues including the brain heart and lungs showing its importance in multiple physiological contexts. Phosphorylation of AKT1 at serine 473 denoted as p-AKT S473 is an important modification that regulates its activity.
Biological function summary

AKT1 regulates a broad spectrum of cellular functions through signaling pathways. It is an important player in the phosphatidylinositol 3-kinase (PI3K) and AKT pathway forming part of a complex involved in promoting cell survival and growth. AKT1 interacts closely with other proteins such as mTOR influencing cellular metabolism and autophagy. The phosphorylation state of AKT1 is critical for its activity with modifications like p-AKT S473 impacting its interaction with cellular substrates.

Pathways

AKT1 participates in the PI3K/AKT/mTOR signaling cascade instrumental in cell proliferation and metabolism. AKT1 integrates signals from insulin and growth factors modulating pathways that control cell growth and glucose uptake. This pathway involves proteins like mTOR and phospho-AKT which coordinate cellular responses to external stimuli. AKT1’s phosphorylation at serine 473 and the involvement of AKT1 E17K a gain-of-function mutation influence these pathways significantly.

AKT1 has associations with cancer and metabolic disorders. Dysregulation of the PI3K/AKT/mTOR pathway involving AKT1 and mTOR often results in oncogenic transformation and uncontrolled cellular proliferation. In cancer such as breast cancer AKT1 mutations including AKT1 E17K are implicated altering cell signals for growth. Additionally AKT1 is connected to metabolic disorders such as Type 2 diabetes where it affects insulin signaling and glucose metabolism highlighting its interaction with metabolic proteins like mTOR and phospho-AKT.
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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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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

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