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PI 3 Kinase p85 beta Recombinant Rabbit Monoclonal Antibody [ST04-77]

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Specification

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Catalog# ET1609-30

PI 3 Kinase p85 beta Recombinant Rabbit Monoclonal Antibody [ST04-77]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PI 3 Kinase p85 beta Recombinant Rabbit Monoclonal Antibody [ST04-77]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human PI 3 Kinase p85 beta aa 136-185 / 728.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC, IP

Molecular Weight

Predicted band size: 82 kDa

Positive Control

Rat heart tissue lysate, PC-12 cell lysate, HeLa cell lysate, NIH/3T3 cell lysates, Hela, SHG-44, human colon tissue, human testis tissue, mouse colon tissue, mouse testis tissue, rat colon tissue, rat testis tissue, HeLa.

Conjugation

unconjugated

Clone Number

ST04-77

RRID

AB_3069841

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Phosphatidylinositol 3-kinase (PI 3-kinase) is composed of p85 and p110 subunits. p85 lacks PI 3-kinase activity and acts as an adapter, coupling p110 to activated protein tyrosine kinase. Two forms of p85 have been described (p85α and p85β), each possessing one SH3 and two SH2 domains. Various p110 isoforms have been identified. p110α and p110β interact with p85α, and p110α has also been shown to interact with p85β in vitro. p110α expression is restricted to white blood cells. It has been shown to bind p85α and p85β, but it apparently does not phosphorylate these subunits. p110α seems to have the capacity to autophosphorylate. p110α does not interact with the p85 subunits. It has been shown to be activated by α and β heterotrimeric G proteins.

Background References

1. Vaca Jacome A.S., et al. N-terminome analysis of the human mitochondrial proteome. Proteomics 15:2519-2524(2015).

2. Mirzaa G.M., et al. Characterisation of mutations of the phosphoinositide-3-kinase regulatory subunit, PIK3R2, in perisylvian polymicrogyria: a next-generation sequencing study. Lancet Neurol. 14:1182-1195(2015).

Sequence Similarity

Belongs to the PI3K p85 subunit family.

Post-translational Modification

Phosphorylated in response to signaling from activated receptor-type protein kinases. Dephosphorylated by PTPRJ. Dephosphorylated at Tyr-655 by PTPN13. Phosphorylation of Tyr-655 impairs while its dephosphorylation promotes interaction with FBXL2 and SCF(FBXL2)-mediated polyubiquitination.; Ubiquitinated. Polyubiquitination by the SCF(FBXL2) complex probably promotes proteasomal degradation of PIK3R2.

Subcellular Location

Cytosol, nucleus

UNIPROT

SwissProt: O00459 Human

SwissProt: O08908 Mouse

SwissProt: Q63788 Rat

Synonyms

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

Expand

Images

  • Western blot analysis of PI 3 Kinase p85 beta on different lysates with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution.<br /><br />Lane 1: Rat heart tissue lysate<br />Lane 2: PC-12 cell lysate<br />Lane 3: HeLa cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 82 kDa<br /><br />Exposure time: Lane 1-2: 3 minutes; Lane 3: 1 minute 2 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PI 3 Kinase p85 beta on different lysates with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

    Lane 1: Rat heart tissue lysate
    Lane 2: PC-12 cell lysate
    Lane 3: HeLa cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 82 kDa

    Exposure time: Lane 1-2: 3 minutes; Lane 3: 1 minute 2 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-30) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of PI 3 Kinase p85 beta on NIH/3T3 cell lysates with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 82 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PI 3 Kinase p85 beta on NIH/3T3 cell lysates with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 82 kDa

    Exposure time: 3 minutes; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-30) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of PI 3 Kinase p85 beta in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of PI 3 Kinase p85 beta in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of PI 3 Kinase p85 beta in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of PI 3 Kinase p85 beta in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-30) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-30) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-30) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling PI 3 Kinase p85 beta.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling PI 3 Kinase p85 beta.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-30, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • PI 3 Kinase p85 beta was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1609-30" style="font-weight: bold;text-decoration: underline;">ET1609-30</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 3 minutes; ECL: K1801

    PI 3 Kinase p85 beta was immunoprecipitated from 0.2 mg HeLa cell lysate with ET1609-30 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1609-30 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: ET1609-30 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of ET1609-30 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 3 minutes; ECL: K1801

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