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Signal Transduction Protein Phosphorylation Ser / Thr Phosphatases

Human PPP2R1B knockout HEK-293T cell lysate (ab258133)

Human PPP2R1B knockout HEK-293T cell lysate (ab258133)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

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Overview

  • Product name

    Human PPP2R1B knockout HEK-293T cell lysate
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 37 bp insertion in exon 1.
  • Passage number

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    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 - found here. 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.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    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.

    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.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab260471 - Human PPP2R1B knockout HEK293T cell lysate 1 x 100µg
    ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Phosphatases
  • Cell type

    epithelial
  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

Target

  • Function

    The PR65 subunit of protein phosphatase 2A serves as a scaffolding molecule to coordinate the assembly of the catalytic subunit and a variable regulatory B subunit.
  • Sequence similarities

    Belongs to the phosphatase 2A regulatory subunit A family.
    Contains 15 HEAT repeats.
  • Domain

    Each HEAT repeat appears to consist of two alpha helices joined by a hydrophilic region, the intrarepeat loop. The repeat units may be arranged laterally to form a rod-like structure.
  • Target information above from: UniProt accession P30154 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • 2AAB_HUMAN
    • beta isoform of regulatory subunit A, protein phosphatase 2
    • PP2A A beta
    • PP2A subunit A isoform PR65-beta
    • PP2A subunit A isoform R1-beta
    • PP2A, subunit A, PR65 beta isoform
    • PP2A, subunit A, R1 beta isoform
    • PPP2R1B
    • PPP4R1
    • PR65B
    • protein phosphatase 2 (formerly 2A), regulatory subunit A (PR 65), beta isoform
    • protein phosphatase 2 (formerly 2A), regulatory subunit A, beta isoform
    • Protein phosphatase 2, regulatory subunit A, beta
    • Protein phosphatase 2, structural/regulatory subunit A, beta
    • protein phosphatase 2, structural/regulatory subunit A, beta isoform
    • serine/threonine protein phosphatase 2A 65 kDa regulatory subunit A, beta isoform
    • serine/threonine protein phosphatase 2A, 65 kDa regulatory subunit A, beta isoform
    • Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A beta isoform
    see all

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab260471 - Human PPP2R1B knockout HEK293T cell lysate 1 x 100µg
    ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Phosphatases
  • Cell type

    epithelial
  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

Images

  • Western blot - Human PPP2R1B knockout HEK293T cell lysate (ab258133)
    Western blot - Human PPP2R1B knockout HEK293T cell lysate (ab258133)

    Lane 1:Wild-type HEK293T cell lysate (20 ug)
    Lane 2:PPP2R1B knockout HEK293T cell lysate (20 ug)
    Lane 3:Caco-2 cell lysate (20 ug)
    Lane 4:HeLa cell lysate (20 ug)

    ab154815 was shown to specifically react with PPP2R1B in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266729 (knockout cell lysate ab258133) was used. Wild-type and PPP2R1B knockout samples were subjected to SDS-PAGE. ab154815 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.

  • Sanger Sequencing - Human PPP2R1B knockout HEK293T cell lysate (ab258133)
    Sanger Sequencing - Human PPP2R1B knockout HEK293T cell lysate (ab258133)
    Homozygous: 37 bp insertion in exon 1

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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