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AB257029

Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate

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PIK3R1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon10.
4 Images
Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)
  • WB

Lab

Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)

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

Lane 2 : PIK3R1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab191606 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

ab191606 Anti-PI 3 Kinase p85 alpha antibody [EPR18702] was shown to specifically react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type and PI 3 Kinase p85 alpha 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. ab191606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-PI 3 Kinase p85 alpha antibody [EPR18702] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-alpha-antibody-epr18702-ab191606'>ab191606</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PIK3R1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pik3r1-pi-3-kinase-p85-alpha-knockout-hela-cell-line-ab265116'>ab265116</a>)

Predicted band size: 83 kDa

Observed band size: 90 kDa

false

Sanger Sequencing - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)
  • Sanger seq

Unknown

Sanger Sequencing - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)

Allele-1 : Insertion of the selection cassette in exon10

Sanger Sequencing - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)
  • Sanger seq

Unknown

Sanger Sequencing - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)

Allele-2 : Insertion of the selection cassette in exon10

Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)
  • WB

Lab

Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (AB257029)

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

Lane 2 : PIK3R1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133595 Recombinant Anti-PI 3 Kinase p85 alpha antibody [EPR5513] was shown to specifically react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type and PI 3 Kinase p85 alpha 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. ab133595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-PI 3 Kinase p85 alpha antibody [EPR5513] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-alpha-antibody-epr5513-ab133595'>ab133595</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (ab257029) at 20 µg

Predicted band size: 83 kDa

Observed band size: 90 kDa

false

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, Insertion of the selection cassette in exon10.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB191606

Anti-PI 3 Kinase p85 alpha antibody [EPR18702]

20ul selling size|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (AB191606)

  • 5
  • 2
Proteins & Peptides

AB84769

Recombinant Human PI 3 Kinase p85 alpha protein (His tag N-Terminus)

Western blot - Recombinant Human PI 3 Kinase p85 alpha protein (His tag N-Terminus) (AB84769)

  • 0
  • 0
Multiplex Kits

AB283852

PI3K/AKT signalling pathway panel

Recombinant

Immunocytochemistry - PI3K/AKT signalling pathway panel (AB283852)

  • 5
  • 1
Primary Antibodies

AB40776

Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y]

20ul selling size|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (AB40776)

  • 0
  • 1
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Reactivity data

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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":"ab257029-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257029-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human PIK3R1 knockout HeLa cell lysate", "number":"AB257029-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
PIK3R1
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.

PI 3 Kinase p85 alpha also known as PI3KR1 or phospho PI3K is a regulatory subunit of phosphoinositide 3-kinase (PI3K) that plays a fundamental role in various signaling pathways. The p85 protein has a molecular weight of approximately 85 kDa. This protein is widely expressed in many cell types facilitating essential cellular functions by forming a heterodimer with the class I PI3K catalytic subunit often implicated in regulating lipid signaling pathways.
Biological function summary

P85 alpha modulates intracellular signaling by interacting with other proteins in a complex formation with catalytic subunit p110. This interaction enables the initiation of downstream signaling cascades controlling cell growth proliferation and survival. PI 3 kinase activity mediated by p85 alpha is integral to cellular homeostasis and metabolism helping to execute cellular responses to a variety of growth factors and hormones.

Pathways

PI 3 Kinase p85 alpha integrates into two principal signaling networks: the PI3K-Akt and mTOR pathways. These pathways are instrumental for regulating anabolic processes and cell survival. The p85 form of PI3K interacts with additional proteins such as Akt and mTOR coordinating cellular energy balance promoting protein synthesis and inhibiting apoptosis. Through these interactions the protein ensures that cells respond accurately to nutritional and stress signals.

Alterations in PI 3 Kinase p85 alpha function have been linked to cancer and type 2 diabetes. Aberrant signaling involving PI3K-Akt-mTOR pathways due to mutations in PI3KR1 can contribute to tumor progression and chemoresistance. Moreover dysfunction of this pathway affects insulin signaling connecting p85 alpha to diabetic pathologies. Proteins such as IRS-1 (insulin receptor substrate 1) act alongside p85 alpha in mediating these disease-associated processes emphasizing its importance in maintaining physiological balance.
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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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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