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PI 3 Kinase p85 alpha Recombinant Rabbit Monoclonal Antibody [SU04-07]

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Specification

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Catalog# ET1608-70

PI 3 Kinase p85 alpha Recombinant Rabbit Monoclonal Antibody [SU04-07]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

PI 3 Kinase p85 alpha Recombinant Rabbit Monoclonal Antibody [SU04-07]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human PI 3 Kinase p85 alpha.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 84 kDa

Positive Control

HepG2 cell lysate, Jurkat cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Hela, MCF-7, HepG2, NIH/3T3, human brain tissue, human lung tissue, mouse lung tissue, rat lung tissue.

Conjugation

unconjugated

Clone Number

SU04-07

RRID

AB_3069823

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:1,000

  • FC

  • 1:50-1:100

Target

Function

Phosphatidylinositol 3-kinase (PI 3-kinase) phosphorylates the 3' OH position of the inositol ring of inositol lipids and is composed of p85 and p110 subunits. PI 3-kinase 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. PI 3-kinase p85α, also known as GRB1, phosphatidylinositol 3-kinase regulatory 1 or p85, is a 724 amino acid protein that exists as four alternatively spliced isoforms. Involved in insulin metabolism, defects in the PI 3-kinase p85α gene have been linked to insulin resistance. PI 3-kinase p85α is polyubiquitinated in T-cells by Cbl-b, and has multiple phosphorylated amino acid residues, including a phosphorylated tyrosine residue at position 467.

Background References

1. Yan LX et al. PIK3R1 targeting by miR-21 suppresses tumor cell migration and invasion by reducing PI3K/AKT signaling and reversing EMT, and predicts clinical outcome of breast cancer. Int J Oncol 48:471-84 (2016).

2. Hu J et al. Filamin B regulates chondrocyte proliferation and differentiation through Cdk1 signaling. PLoS One 9:e89352 (2014).

Sequence Similarity

Belongs to the PI3K p85 subunit family.

Tissue Specificity

Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level).

Post-translational Modification

Polyubiquitinated in T-cells by CBLB; which does not promote proteasomal degradation but impairs association with CD28 and CD3Z upon T-cell activation.; Phosphorylated. Tyrosine phosphorylated in response to signaling by FGFR1, FGFR2, FGFR3 and FGFR4. Phosphorylated by CSF1R. Phosphorylated by ERBB4. Phosphorylated on tyrosine residues by TEK/TIE2. Dephosphorylated by PTPRJ. Phosphorylated by PIK3CA at Ser-608; phosphorylation is stimulated by insulin and PDGF. The relevance of phosphorylation by PIK3CA is however unclear (By similarity). Phosphorylated in response to KIT and KITLG/SCF. Phosphorylated by FGR.

Subcellular Location

Cytoplasm, membrane, nucleus.

UNIPROT

SwissProt: P27986 Human

SwissProt: P26450 Mouse

SwissProt: Q63787 Rat

Synonyms

GRB1 antibody

p85 alpha antibody

p85 antibody

P85A_HUMAN antibody

Phosphatidylinositol 3 kinase associated p 85 alpha antibody

Phosphatidylinositol 3 kinase regulatory 1 antibody

Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 1 (p85 alpha) antibody

Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha antibody

Phosphatidylinositol 3-kinase regulatory subunit alpha antibody

Phosphoinositide 3 kinase, regulatory subunit 1 (alpha) antibody

Expand

Images

  • Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Rabbit anti-PI 3 Kinase p85 alpha antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>) at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: Jurkat cell lysate (20 µg/Lane)<br />Lane 3: MCF7 cell lysate (20 µg/Lane)<br />Lane 4: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 5: C6 cell lysate (20 µg/Lane)<br />Lane 6: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 7: Rat brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 kDa<br /><br />Exposure time: 1 minute; 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/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) 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 alpha on different lysates with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: Jurkat cell lysate (20 µg/Lane)
    Lane 3: MCF7 cell lysate (20 µg/Lane)
    Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 5: C6 cell lysate (20 µg/Lane)
    Lane 6: Mouse brain tissue lysate (40 µg/Lane)
    Lane 7: Rat brain tissue lysate (40 µg/Lane)

    Predicted band size: 84 kDa
    Observed band size: 84 kDa

    Exposure time: 1 minute; 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 (ET1608-70) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. 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 alpha in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 alpha in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 alpha in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 alpha in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 alpha in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 alpha in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 alpha in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;594 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 alpha in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 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 brain tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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 brain tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) 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 (ET1608-70) 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 lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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 lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) 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 (ET1608-70) 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 lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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 lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) 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 (ET1608-70) 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 lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</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 lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) 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 (ET1608-70) 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 PI 3 Kinase p85 alpha was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1608-70" style="font-weight: bold;text-decoration: underline;">ET1608-70</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of PI 3 Kinase p85 alpha was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-70, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation

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