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AB270499

Human PPP2R5D knockout A-431 cell lysate

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PPP2R5D KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 7 bp deletion Frameshift = 99.91%.

View Alternative Names

2A5D_HUMAN, B'delta, B56D, Delta isoform of regulatory subunit B56, protein phosphatase 2A, MGC2134, MGC8949, OTTHUMP00000039821, PP2A B subunit B' delta isoform, PP2A B subunit B56 delta isoform, PP2A B subunit PR61 delta isoform, PP2A B subunit R5 delta isoform, PP2A B subunit isoform B''-delta, PP2A B subunit isoform B56-delta, PP2A B subunit isoform PR61-delta, PP2A B subunit isoform R5-delta, Protein phosphatase 2 regulatory subunit B (B56) delta isoform, Protein phosphatase 2 regulatory subunit B delta isoform, Protein phosphatase 2 regulatory subunit B' delta, Serine threonine protein phosphatase 2A 56 kDa regulatory subunit delta isoform, Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform, TEG-271, Tex271

3 Images
Western blot - Human PPP2R5D knockout A-431 cell lysate (AB270499)
  • WB

Lab

Western blot - Human PPP2R5D knockout A-431 cell lysate (AB270499)

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : PPP2R5D knockout A431 cell lysate 20 ug

Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab188323 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab188323 was shown to react with PPP2R5D in wild-type A-431 cells in western blot with loss of signal observed in PPP2R5D knockout cell line ab270476 (knockout cell lysate ab270499). Wild-type and PPP2R5D knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188323 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PPP2R5D antibody [EPR15617] (<a href='/en-us/products/primary-antibodies/ppp2r5d-antibody-epr15617-ab188323'>ab188323</a>) at 1/10000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

PPP2R5D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PPP2R5D knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-ppp2r5d-knockout-a-431-cell-line-ab270476'>ab270476</a>)

Lane 3:

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

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 70 kDa

Observed band size: 60-65 kDa

false

Western blot - Human PPP2R5D knockout A-431 cell lysate (AB270499)
  • WB

Lab

Western blot - Human PPP2R5D knockout A-431 cell lysate (AB270499)

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : PPP2R5D knockout A431 cell lysate 20 ug

Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab188325 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab188325 was shown to react with PPP2R5D in wild-type A-431 cells in western blot with loss of signal observed in PPP2R5D knockout cell line ab270476 (knockout cell lysate ab270499). Wild-type and PPP2R5D knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188325 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PPP2R5D antibody [EPR15617-50] (<a href='/en-us/products/primary-antibodies/ppp2r5d-antibody-epr15617-50-ab188325'>ab188325</a>) at 1/10000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

PPP2R5D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PPP2R5D knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-ppp2r5d-knockout-a-431-cell-line-ab270476'>ab270476</a>)

Lane 3:

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

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 70 kDa

Observed band size: 60-65 kDa

false

Next Generation Sequencing - Human PPP2R5D knockout A-431 cell lysate (AB270499)
  • NGS

Supplier Data

Next Generation Sequencing - Human PPP2R5D knockout A-431 cell lysate (AB270499)

Knockout achieved by CRISPR/Cas9; X = 7 bp deletion; Frameshift = 99.91%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 7 bp deletion Frameshift = 99.91%

Disease

Epidermoid Carcinoma

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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 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
PPP2R5D
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.

PPP2R5D also known as B56δ forms part of the regulatory subunit of protein phosphatase 2A (PP2A). Protein phosphatase 2A is an important enzyme in cellular signaling that dephosphorylates target proteins. The approximate molecular mass of PPP2R5D is 72 kDa. It is found expressed in various tissues throughout the human body but its levels can vary between different cell types.
Biological function summary

PPP2R5D plays an essential role in the regulation of cell growth and division. This protein is part of the PP2A holoenzyme complex which assembles with catalytic and scaffolding subunits. Through its regulatory influence PPP2R5D affects the phosphorylation state of several substrates thereby impacting cellular processes like cell cycle progression and apoptosis.

Pathways

PPP2R5D significantly contributes to the MAPK signaling pathway and the Wnt pathway. It modulates phosphorylation states affecting signal transduction mechanisms. In the MAPK pathway PPP2R5D interacts with proteins like ERK1/2 by influencing their activity. Similarly in the Wnt pathway it acts on components important for maintaining cellular balance and signaling fidelity.

PPP2R5D mutations and dysregulation have been linked to intellectual disabilities and cancer. Changes in its function are associated with Autism Spectrum Disorder where it potentially interacts with other proteins involved in neuronal development. In cancer altered PPP2R5D activity may disrupt normal cellular growth control often involving proteins like c-Myc that are critical for proliferation and survival.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB188323

Anti-PPP2R5D antibody [EPR15617]

RabMAb|Recombinant|KO Validated|Trial Size|Trial Size

Immunocytochemistry/ Immunofluorescence - Anti-PPP2R5D antibody [EPR15617] (AB188323)

  • 0
  • 0
Proteins & Peptides

AB114547

Recombinant Human PPP2R5D protein

SDS-PAGE - Recombinant Human PPP2R5D protein (AB114547)

  • 0
  • 0
Cell Lines & Lysates

AB270476

Human PPP2R5D knockout A-431 cell line

Advanced Validation

Western blot - Human PPP2R5D knockout A-431 cell line (AB270476)

  • 0
  • 0

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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