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AB257824

Human AK1 knockout HeLa cell lysate

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AK1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon4 and 1 bp deletion in exon4 and 5 bp deletion in exon4.
4 Images
Western blot - Human AK1 knockout HeLa cell lysate (AB257824)
  • WB

Unknown

Western blot - Human AK1 knockout HeLa cell lysate (AB257824)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : AK1 knockout HeLa cell lysate (20 ug)
Lane 3 : U-87MG cell lysate (20 ug)

ab138504 was shown to specifically react with AK 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265132 (knockout cell lysate ab257824) was used. Wild-type and AK 1 knockout samples were subjected to SDS-PAGE. ab138504 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-AK 1 antibody [EP7083] (<a href='/en-us/products/primary-antibodies/ak-1-antibody-ep7083-ab138504'>ab138504</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

AK1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human AK1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ak1-knockout-hela-cell-line-ab265132'>ab265132</a>)

Lane 3:

U-87MG 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: 22 kDa

Observed band size: 25 kDa

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Sanger Sequencing - Human AK1 knockout HeLa cell lysate (AB257824)
  • Sanger seq

Unknown

Sanger Sequencing - Human AK1 knockout HeLa cell lysate (AB257824)

Allele-3 : 1 bp deletion in exon4

Sanger Sequencing - Human AK1 knockout HeLa cell lysate (AB257824)
  • Sanger seq

Unknown

Sanger Sequencing - Human AK1 knockout HeLa cell lysate (AB257824)

Allele-1 : 14 bp deletion in exon4

Sanger Sequencing - Human AK1 knockout HeLa cell lysate (AB257824)
  • Sanger seq

Unknown

Sanger Sequencing - Human AK1 knockout HeLa cell lysate (AB257824)

Allele-2 : 5 bp deletion in exon4

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, 14 bp deletion in exon4 and 1 bp deletion in exon4 and 5 bp deletion in exon4.

Disease

Adenocarcinoma

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Primary Antibodies

AB138504

Anti-AK 1 antibody [EP7083]

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Western blot - Anti-AK 1 antibody [EP7083] (AB138504)

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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 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
AK1
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.

Adenylate kinase 1 also known as AK1 plays a critical role in cellular energy homeostasis. It catalyzes the reversible transfer of a phosphate group from ATP to AMP forming two ADP molecules. AK1 is a 24-kilodalton protein largely expressed in tissues with high-energy demands like brain heart and skeletal muscle. Expression levels vary but AK1's presence is notably higher in these energy-intense regions highlighting its importance in energy transfer and equilibrium.
Biological function summary

Adenylate kinase 1 is essential for maintaining the adenine nucleotide pool balance. It does not act alone but functions within diverse cellular contexts ensuring efficient energy supply which is necessary for critical cellular processes. In these contexts AK1 forms part of a larger network that involves other kinases and molecules operating to sustain sufficient energy levels during periods of high activity or stress. This orchestration of molecules ensures cells adapt to changing energy requirements swiftly.

Pathways

AK1 is a critical component of the ATP-generating phosphotransfer network and is involved in the cellular adenylate kinase reaction pathway. It interacts closely with proteins such as AMP-activated protein kinase (AMPK) which assists in detecting and responding to energy deficits. In the context of the phosphotransfer network maintaining the balance between ATP ADP and AMP is important and AK1 enables this dynamic equilibrium. By interrelating with these pathways AK1 supports efficient energy transduction and cellular adaptation to metabolic changes.

Abnormal AK1 function or expression has been implicated in metabolic conditions like mitochondrial myopathy and heart diseases such as myocardial infarction. A disruption in the balance of adenylate kinase activity can lead to energy deficiency aggravating these conditions. Connections to these diseases involve proteins such as creatine kinase which plays a role in energy buffering. The interplay between AK1 and associated kinases is key in understanding how energetic imbalances contribute to the development and progression of 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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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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