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AB259055

Human PPM1A knockout HeLa cell lysate

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PPM1A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon3 and Insertion of the selection cassette in exon3.
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Western blot - Human PPM1A knockout HeLa cell lysate (AB259055)
  • WB

Lab

Western blot - Human PPM1A knockout HeLa cell lysate (AB259055)

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 2 : PPM1A knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 3 : HAP1 whole cell lyate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

ab173527 was shown to specifically react with GNAI3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265348 (knockout cell lysate ab259055) was used. Wild-type and GNAI3 knockout samples were subjected to SDS-PAGE. ab173527 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4oC at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PPM1A antibody [p6c7] (<a href='/en-us/products/primary-antibodies/ppm1a-antibody-p6c7-ab14824'>ab14824</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

PPM1A knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PPM1A knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ppm1a-knockout-hela-cell-line-ab265348'>ab265348</a>)

Lane 3:

HAP1 whole cell lyate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 41 kDa,42 kDa

Observed band size: 42 kDa

false

Sanger Sequencing - Human PPM1A knockout HeLa cell lysate (AB259055)
  • Sanger seq

Unknown

Sanger Sequencing - Human PPM1A knockout HeLa cell lysate (AB259055)

Allele-1 : 4 bp deletion in exon3

Sanger Sequencing - Human PPM1A knockout HeLa cell lysate (AB259055)
  • Sanger seq

Unknown

Sanger Sequencing - Human PPM1A knockout HeLa cell lysate (AB259055)

Allele-2 : Insertion of the selection cassette in exon3

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, 4 bp deletion in exon3 and Insertion of the selection cassette in exon3.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB14824

Anti-PPM1A antibody [p6c7]

KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1A antibody [p6c7] (AB14824)

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Proteins & Peptides

AB51205

Recombinant human PPM1A protein

Bioactive

SDS-PAGE - Recombinant human PPM1A protein (AB51205)

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Cell Lines & Lysates

AB265348

Human PPM1A knockout HeLa cell line

Advanced Validation

Western blot - Human PPM1A knockout HeLa cell line (AB265348)

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  • 0
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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":"ab259055-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human PPM1A knockout HeLa cell lysate", "number":"AB259055-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB259055-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
PPM1A
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.

PPM1A also known as protein phosphatase 1A or PP2C-alpha functions as a serine/threonine phosphatase. This protein plays an important role in removing phosphate groups from serine and threonine residues on proteins. The molecular mass of PPM1A is approximately 42-44 kDa. Expression of PPM1A is widespread and found in various human tissues indicating its significant involvement in multiple cellular processes.
Biological function summary

PPM1A regulates many cellular mechanisms by dephosphorylating targets and acting as a negative regulator of several signaling pathways. It modulates stress responses cell cycle progression and immune responses. PPM1A does not typically form complexes with other proteins but interacts closely with specific signaling molecules to control their activity. This regulation influences cellular homeostasis and growth in response to external and internal signals.

Pathways

PPM1A influences the TGF-beta signaling pathway where it interacts with and deactivates SMAD proteins by dephosphorylation. It also participates in the p38 MAPK pathway. These interactions highlight its role in controlling transduction signals initiated by growth factors and stress. By regulating these pathways PPM1A maintains a balance in cellular responses to growth and environmental stresses.

PPM1A links to cancer and inflammatory diseases. Its ability to modulate phosphorylation states impacts cell proliferation and immune responses critical factors in these conditions. For instance PPM1A interaction with SMAD3 in the TGF-beta pathway can influence tumor progression. Additionally its regulation of proteins such as p38 MAPK can alter inflammatory responses affecting inflammatory disease severity.
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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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