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

Usd: 385

Specification

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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

Overview

Product Name

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

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

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

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 84 kDa/55 kDa

Positive Control

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.

Conjugation

unconjugated

Clone Number

PSH01-38

RRID

AB_3072785

Reactivity Data

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

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

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

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.

Background References

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

Subcellular Location

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

UNIPROT

SwissProt: P27986 Human

SwissProt: P26450 Mouse

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

  • <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.

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

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