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PI 3 Kinase p85 alpha Mouse Monoclonal Antibody [L3-D6]

Usd: 350

Specification

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Catalog# EM1701-62

PI 3 Kinase p85 alpha Mouse Monoclonal Antibody [L3-D6]

Application

  • WB

  • IHC-P

  • IF-Cell

  • IF-Tissue

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 alpha Mouse Monoclonal Antibody [L3-D6]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human PI3-kinase p85 subunit alpha aa 19-219 / 724.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 84 kDa

Positive Control

MCF7 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, Mouse brain tissue lysate, Rat liver tissue lysate, Raji cell lysate, C6 cell lysate, HepG2, LOVO, human brain tissue, human lung tissue, mouse lung tissue, rat lung tissue.

Conjugation

unconjugated

Clone Number

L3-D6

RRID

AB_3068786

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*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:1,000

  • IF-Tissue

  • 1:200

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. p110δ expression is restricted to white blood cells. It has been shown to bind p85α and β, but it apparently does not phosphorylate these subunits. p110δ seems to have the capacity to autophosphorylate. p110γ does not interact with the p85 subunits. It has been shown to be activated by α and βγ heterotrimeric G proteins.

Background References

1. Vainikka S et al. Signal transduction by fibroblast growth factor receptor-4 (FGFR-4). Comparison with FGFR-1. J Biol Chem 269:18320-18326 (1994).

2. Winnay J N et al. A regulatory subunit of phosphoinositide 3-kinase increases the nuclear accumulation of X-box-binding protein-1 to modulate the unfolded protein response. Nat Med 16:438-445 (2010).

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

Cytosol. Endoplasmic reticulum. Golgi apparatus. Nucleus. Plasma Membrane.

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 Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7 cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 4: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 5: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 kDa<br /><br />Exposure time: 3 minutes; 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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</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 Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) at 1/1,000 dilution.

    Lane 1: MCF7 cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: RAW264.7 cell lysate (20 µg/Lane)
    Lane 4: Mouse brain tissue lysate (40 µg/Lane)
    Lane 5: Rat liver tissue lysate (40 µg/Lane)

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

    Exposure time: 3 minutes; 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 (EM1701-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) 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 Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</a>) at 1/1,000 dilution.<br /><br />Lane 1: Raji cell lysate<br />Lane 2: NIH/3T3 cell lysate<br />Lane 3: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 kDa<br /><br />Exposure time: 2 minutes; 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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</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 Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) at 1/1,000 dilution.

    Lane 1: Raji cell lysate
    Lane 2: NIH/3T3 cell lysate
    Lane 3: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 2 minutes; 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 (EM1701-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</a>) at 1/1,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si PI 3 Kinase p85 alpha cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) at 1/1,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si PI 3 Kinase p85 alpha cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 84 kDa
    Observed band size: 84 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 (EM1701-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining PI 3 Kinase p85 alpha (green) in HepG2 cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining PI 3 Kinase p85 alpha (green) in HepG2 cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • ICC staining PI 3 Kinase p85 alpha (green) in LOVO cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining PI 3 Kinase p85 alpha (green) in LOVO cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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 Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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 Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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 Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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 Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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 Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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 Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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/EM1701-62" style="font-weight: bold;text-decoration: underline;">EM1701-62</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 Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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