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PI 3 Kinase p85 alpha Recombinant Mouse Monoclonal Antibody [A3-D0-R]

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Catalog# HA601206

PI 3 Kinase p85 alpha Recombinant Mouse Monoclonal Antibody [A3-D0-R]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

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

概述

产品名称

PI 3 Kinase p85 alpha Recombinant Mouse Monoclonal Antibody [A3-D0-R]

抗体类型

Recombinant Mouse Monoclonal Antibody

免疫原

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

种属反应性

Human

验证应用

WB, IF-Cell

分子量

Predicted band size: 84 kDa

阳性对照

SW480 cell lysate, A431 cell lysate, 293T cell lysate, Jurkat cell lysate, Raji cell lysate, MCF7 cell lysate, A549 cell lysate, U-937.

偶联

unconjugated

克隆号

A3-D0-R

RRID

AB_3071925

产品特性

形态

Liquid

浓度

1ug/ul

存放说明

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

存储缓冲液

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG1

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:100

发表文章中的应用

WB See 3 publications below

靶点

功能

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 γ, 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.

背景文献

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

2. Schmidt JW et al. Stat5 regulates the phosphatidylinositol 3-kinase/Akt1 pathway during mammary gland development and tumorigenesis. Mol Cell Biol 34:1363-77 (2014).

亚细胞定位

Cytosol, Membrane, Nucleus.

UNIPROT #

SwissProt: P27986 Human

别名

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

展开

图片

  • Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: SW480 cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: 293T cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: Raji cell lysate<br />Lane 6: MCF7 cell lysate<br />Lane 7: A549 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 kDa<br /><br />Exposure time: 1 minute 40 seconds; ECL: K1802;<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/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 (HA601206) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: SW480 cell lysate
    Lane 2: A431 cell lysate
    Lane 3: 293T cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: Raji cell lysate
    Lane 6: MCF7 cell lysate
    Lane 7: A549 cell lysate

    Lysates/proteins at 15 µg/Lane.

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

    Exposure time: 1 minute 40 seconds; 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 (HA601206) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

  • Immunocytochemistry analysis of U-937 cells labeling PI 3 Kinase p85 alpha with Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of U-937 cells labeling PI 3 Kinase p85 alpha with Mouse anti-PI 3 Kinase p85 alpha antibody (HA601206) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PI 3 Kinase p85 alpha antibody (HA601206) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (<a href="/products/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/1,000 dilution.<br /><br />Lane 1: SW480 cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: 293T cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: A549 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: 1 minute 20 seconds;<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/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</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 (HA601206) at 1/1,000 dilution.

    Lane 1: SW480 cell lysate
    Lane 2: A431 cell lysate
    Lane 3: 293T cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: A549 cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 1 minute 20 seconds;

    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 (HA601206) 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/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/5,000 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: 2 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/HA601206" style="font-weight: bold;text-decoration: underline;">HA601206</a>) at 1/5,000 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 (HA601206) at 1/5,000 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: 2 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 (HA601206) at 1/5,000 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.

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

引文

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  1. WB
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