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Signal Transduction Signaling Pathway Lipid Signaling Lipid Kinases
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Validated using a knockout cell line

Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)

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  • SDS
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Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)
  • Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)

Key features and details

  • Mouse monoclonal [T15] to PI 3 Kinase p85 beta
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-PI 3 Kinase p85 beta antibody [T15]
    See all PI 3 Kinase p85 beta primary antibodies
  • Description

    Mouse monoclonal [T15] to PI 3 Kinase p85 beta
  • Host species

    Mouse
  • Specificity

    T15 recognises the p85 beta subunit of bovine PI 3 kinase. It does not cross react with the p85 alpha subunit.
  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data
  • Immunogen

    Balculovirus expressed (Recombinant) bovine P85 beta subunit of PI 3 kinase.

  • Positive control

    • WB: HEK293T cell lysate; Lysates of SF9 cells infected with baculovirus expressing p85 beta.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer

    Preservative: 0.09% Sodium azide
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    This antibody is purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
  • Clonality

    Monoclonal
  • Clone number

    T15
  • Myeloma

    NS1
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Lipid Signaling
    • Lipid Kinases
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • KO cell lines

    • Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (ab266799)
  • KO cell lysates

    • Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell lysate (ab257586)
  • Recombinant Protein

    • Recombinant Human PI 3 Kinase p85 beta protein (ab125568)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab28356 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
WB
Human
All applications
Mouse
Rat
Application Abreviews Notes
WB
1/1000 - 1/5000. Predicted molecular weight: 82 kDa.
Notes
WB
1/1000 - 1/5000. Predicted molecular weight: 82 kDa.

Target

  • Function

    Regulatory subunit of phosphoinositide-3-kinase (PI3K), a kinase that phosphorylates PtdIns(4,5)P2 (Phosphatidylinositol 4,5-bisphosphate) to generate phosphatidylinositol 3,4,5-trisphosphate (PIP3). PIP3 plays a key role by recruiting PH domain-containing proteins to the membrane, including AKT1 and PDPK1, activating signaling cascades involved in cell growth, survival, proliferation, motility and morphology. Binds to activated (phosphorylated) protein-tyrosine kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Indirectly regulates autophagy (PubMed:23604317). Promotes nuclear translocation of XBP1 isoform 2 in a ER stress- and/or insulin-dependent manner during metabolic overloading in the liver and hence plays a role in glucose tolerance improvement.
  • Involvement in disease

    Megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome 1
  • Sequence similarities

    Belongs to the PI3K p85 subunit family.
    Contains 1 Rho-GAP domain.
    Contains 2 SH2 domains.
    Contains 1 SH3 domain.
  • Domain

    The SH2 2 domain is required for interaction with FBXL2 and PTPN13.
  • Post-translational
    modifications

    Phosphorylated in response to signaling from activated receptor-type protein kinases (PubMed:19690332, PubMed:20068231). Dephosphorylated by PTPRJ (PubMed:18348712). Dephosphorylated at Tyr-655 by PTPN13. Phosphorylation of Tyr-655 impairs while its dephosphorylation promotes interaction with FBXL2 and SCF(FBXL2)-mediated polyubiquitination (PubMed:23604317).
    Ubiquitinated. Polyubiquitination by the SCF(FBXL2) complex probably promotes proteasomal degradation of PIK3R2.
  • Target information above from: UniProt accession O00459 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5296 Human
    • Entrez Gene: 18709 Mouse
    • Entrez Gene: 29741 Rat
    • Omim: 603157 Human
    • SwissProt: O00459 Human
    • SwissProt: O08908 Mouse
    • SwissProt: Q63788 Rat
    • Unigene: 371344 Human
    • Unigene: 12945 Mouse
    • Unigene: 22497 Rat
    see all
  • Alternative names

    • p85 antibody
    • p85 beta antibody
    • P85B antibody
    • P85B_HUMAN antibody
    • Phosphatidylinositol 3 kinase antibody
    • Phosphatidylinositol 3 kinase regulatory beta subunit antibody
    • Phosphatidylinositol 3 kinase regulatory subunit beta antibody
    • Phosphatidylinositol 3 kinase regulatory subunit polypeptide 2 antibody
    • Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 2 (p85 beta) antibody
    • Phosphatidylinositol 3-kinase 85 kDa regulatory subunit beta antibody
    • Phosphatidylinositol 3-kinase regulatory subunit beta antibody
    • Phosphoinositide 3 kinase regulatory subunit 2 (beta) antibody
    • Phosphoinositide 3 kinase regulatory subunit 2 antibody
    • Phosphoinositide 3 kinase regulatory subunit polypeptide 2 (p85 beta) antibody
    • Phosphoinositide 3 kinase regulatory subunit polypeptide 2 antibody
    • Phosphoinositide 3 kinase, regulatory subunit 2 (beta) antibody
    • Phosphoinositide 3 kinase, regulatory subunit 2 (p85 beta) antibody
    • PI3 kinase p85 beta subunit antibody
    • PI3 kinase p85 subunit beta antibody
    • PI3-kinase regulatory subunit beta antibody
    • PI3-kinase subunit p85-beta antibody
    • PI3K antibody
    • PI3K regulatory subunit beta antibody
    • PIK3R 2 antibody
    • PIK3R2 antibody
    • PtdIns 3 kinase p85 beta antibody
    • PtdIns-3-kinase regulatory subunit beta antibody
    • PtdIns-3-kinase regulatory subunit p85-beta antibody
    see all

Images

  • Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)
    Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)
    All lanes : Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : PIK3R2 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 82 kDa
    Observed band size: 85 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab28356 observed at 85 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

     ab28356 was shown to react with PI 3 Kinase p85 beta in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266799 (knockout cell lysate ab257586) was used. Wild-type HEK-293T and PIK3R2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab28356 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)
    Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (ab28356)
    Western Blotting of HEK293 and HepG2 cell lysates with Mouse anti Bovine PI-3 kinase p85 subunit beta (ab28356), recombinant p85ß is included as a positive control.

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (10)

Publishing research using ab28356? Please let us know so that we can cite the reference in this datasheet.

ab28356 has been referenced in 10 publications.

  • Zheng D  et al. Long Non-coding RNA Gas5 Is Associated With Preeclampsia and Regulates Biological Behaviors of Trophoblast via MicroRNA-21. Front Genet 11:188 (2020). PubMed: 32194641
  • Pedini F  et al. Joint action of miR-126 and MAPK/PI3K inhibitors against metastatic melanoma. Mol Oncol 13:1836-1854 (2019). PubMed: 31115969
  • Dai K  et al. Kbtbd2 inhibits the cytotoxic activity of immortalized NK cells through down-regulating mTOR signaling in a mouse hepatocellular carcinoma model. Eur J Immunol 48:683-695 (2018). PubMed: 29331106
  • Kuchay S  et al. PTEN counteracts FBXL2 to promote IP3R3- and Ca2+-mediated apoptosis limiting tumour growth. Nature 546:554-558 (2017). PubMed: 28614300
  • Li HB  et al. Long Non-Coding RNA-MALAT1 Mediates Retinal Ganglion Cell Apoptosis Through the PI3K/Akt Signaling Pathway in Rats with Glaucoma. Cell Physiol Biochem 43:2117-2132 (2017). PubMed: 29065394
  • Felli N  et al. AP2a controls the dynamic balance between miR-126&126* and miR-221&222 during melanoma progression. Oncogene 35:3016-26 (2016). WB . PubMed: 26434590
  • Du C  et al. MiR-126-3p suppresses tumor metastasis and angiogenesis of hepatocellular carcinoma by targeting LRP6 and PIK3R2. J Transl Med 12:259 (2014). Human . PubMed: 25240815
  • Ketscher A  et al. LSD1 controls metastasis of androgen-independent prostate cancer cells through PXN and LPAR6. Oncogenesis 3:e120 (2014). WB, IP ; Human . PubMed: 25285406
  • Kuchay S  et al. FBXL2- and PTPL1-mediated degradation of p110-free p85ß regulatory subunit controls the PI(3)K signalling cascade. Nat Cell Biol 15:472-80 (2013). Human . PubMed: 23604317
  • Krishnan S  et al. p85ß regulatory subunit of class IA PI3 kinase negatively regulates mast cell growth, maturation, and leukemogenesis. Blood 119:3951-61 (2012). WB . PubMed: 22378847

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