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Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) Recombinant Rabbit Monoclonal Antibody [PSH01-38]

RMB: 900 特惠 1600

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Catalog# HA721672

Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) Recombinant Rabbit Monoclonal Antibody [PSH01-38]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

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

概述

产品名称

Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) Recombinant Rabbit Monoclonal Antibody [PSH01-38]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within human PIK3R1 aa 451-500 / 724.

种属反应性

Human, Mouse

验证应用

WB, IF-Cell

分子量

Predicted band size: 84 kDa/55 kDa

阳性对照

NIH/3T3 treated with 1mM sodium orthovanadate for 30 minutes cell lysate, NIH/3T3 cells treated with 1mM Sodium orthovanadate for 30 minutes, 293T overexpress Src cell lysate, 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate.

偶联

unconjugated

克隆号

PSH01-38

RRID

AB_3072785

反应性数据

WB IF-Cell
Human
Tested Tested
Mouse
Tested Tested
Tested Tested
Rabbit
Tested
Zebrafish
Tested
Rat
Tested

产品特性

形态

Liquid

浓度

存放说明

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.

存储缓冲液

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

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

靶点

功能

Phosphatidylinositol 3-kinase regulatory subunit alpha is an enzyme that in humans is encoded by the PIK3R1 gene. Phosphatidylinositol 3-kinase phosphorylates the inositol ring of phosphatidylinositol at the 3-prime position. The enzyme comprises a 110 kD catalytic subunit and a regulatory subunit of either 85, 55, or 50 kD. This gene encodes the 85 kD regulatory subunit. Phosphatidylinositol 3-kinase plays an important role in the metabolic actions of insulin, and a mutation in this gene has been associated with insulin resistance. Alternative splicing of this gene results in three transcript variants encoding different isoforms.

背景文献

1. Koch PA et al. The molecular mechanisms mediating class II PI 3-kinase function in cell physiology. FEBS J. 2021 Dec

2. Acosta-Martinez M et al. The PI3K/Akt Pathway in Meta-Inflammation. Int J Mol Sci. 2022 Dec

亚细胞定位

Cell-cell junction, cis-Golgi network, cytoplasm, cytosol, membrane, nucleus.

UNIPROT

SwissProt: P27986 Human

SwissProt: P26450 Mouse

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

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (<a href="/products/HA721672" style="font-weight: bold;text-decoration: underline;">HA721672</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: NIH/3T3 treated with 1mM sodium orthovanadate for 30 minutes cell lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 55/85 kDa<br /><br />Exposure time: 3 minutes 10 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/HA721672" style="font-weight: bold;text-decoration: underline;">HA721672</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 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: NIH/3T3 treated with 1mM sodium orthovanadate for 30 minutes cell lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 84 kDa
    Observed band size: 55/85 kDa

    Exposure time: 3 minutes 10 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 (HA721672) 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 1/100,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of NIH/3T3 cells treated with or without 1mM Sodium orthovanadate for 30 minutes labeling Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (<a href="/products/HA721672" style="font-weight: bold;text-decoration: underline;">HA721672</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 Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (<a href="/products/HA721672" style="font-weight: bold;text-decoration: underline;">HA721672</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of NIH/3T3 cells treated with or without 1mM Sodium orthovanadate for 30 minutes labeling Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) 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 Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (<a href="/products/HA721672" style="font-weight: bold;text-decoration: underline;">HA721672</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T overexpress Src cell lysate<br />Lane 2: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate<br />Lane 3: 293T overexpress Src cell lysate, the membrane treated with λpp for 1 hour<br />Lane 4: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 28 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/HA721672" style="font-weight: bold;text-decoration: underline;">HA721672</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 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) at 1/1,000 dilution.

    Lane 1: 293T overexpress Src cell lysate
    Lane 2: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate
    Lane 3: 293T overexpress Src cell lysate, the membrane treated with λpp for 1 hour
    Lane 4: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 84 kDa
    Observed band size: 55 kDa

    Exposure time: 28 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 (HA721672) 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 1/100,000 dilution was used for 1 hour at room temperature.

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

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