Name: Anti-PI 3 Kinase p85 alpha antibody [EPR18702]
SKU: ab191606
Rating: 5 (2 reviews)

Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) is a rabbit monoclonal antibody detecting PI 3 Kinase p85 alpha in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, ICC/IF. Suitable for African green monkey, Human, Mouse, Rat.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 220 publications

Images

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (AB191606), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Tested	Tested
Rat	Expected	Tested	Expected	Expected
African green monkey	Expected	Tested	Expected	Expected
Recombinant full length protein - Human	Not recommended	Tested	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat, African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species African green monkey	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/250	Notes -
Species Human	Dilution info 1/250	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/150	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Species Human	Dilution info 1/150	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Rat, African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info -	Notes -

Associated Products

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Target data

See full target information PIK3R1

Function

Binds to activated (phosphorylated) protein-Tyr kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Necessary for the insulin-stimulated increase in glucose uptake and glycogen synthesis in insulin-sensitive tissues. Plays an important role in signaling in response to FGFR1, FGFR2, FGFR3, FGFR4, KITLG/SCF, KIT, PDGFRA and PDGFRB. Likewise, plays a role in ITGB2 signaling (PubMed:17626883, PubMed:19805105, PubMed:7518429). Modulates the cellular response to ER stress by promoting nuclear translocation of XBP1 isoform 2 in a ER stress- and/or insulin-dependent manner during metabolic overloading in the liver and hence plays a role in glucose tolerance improvement (PubMed:20348923).

Alternative names

GRB1, PIK3R1, Phosphatidylinositol 3-kinase regulatory subunit alpha, PI3-kinase regulatory subunit alpha, PI3K regulatory subunit alpha, PtdIns-3-kinase regulatory subunit alpha, Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha, PI3-kinase subunit p85-alpha, PtdIns-3-kinase regulatory subunit p85-alpha

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR18702
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunocytochemistry/immunofluorescence (ICC/IF) in African green monkey, Human, Mouse, Rat samples.

What is the molecular weight of PI 3 Kinase p85 alpha?

Anti-PI 3 Kinase p85 alpha [EPR18702] (ab191606) specifically detects a band for PI 3 Kinase p85 alpha (UniProt: Q63787) at a molecular weight of 84kDa.

Trusted by the scientific community

Anti-PI 3 Kinase p85 alpha [EPR18702] (ab191606) was first used in a scientific publication in 2016 and has been cited over 220 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) has been confirmed by Western blot testing in PIK3R1 Knockout HAP1 cells.

Other related products

We have a range of other formats of antibody clone [EPR18702] also available for your convenience:
ab191606, Carrier free - ab223792, APC - ab225369, PE - ab225370, Alexa Fluor® 488 - ab225371

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

PI 3 Kinase p85 alpha also known as PI3KR1 or phospho PI3K is a regulatory subunit of phosphoinositide 3-kinase (PI3K) that plays a fundamental role in various signaling pathways. The p85 protein has a molecular weight of approximately 85 kDa. This protein is widely expressed in many cell types facilitating essential cellular functions by forming a heterodimer with the class I PI3K catalytic subunit often implicated in regulating lipid signaling pathways.

Biological function summary

P85 alpha modulates intracellular signaling by interacting with other proteins in a complex formation with catalytic subunit p110. This interaction enables the initiation of downstream signaling cascades controlling cell growth proliferation and survival. PI 3 kinase activity mediated by p85 alpha is integral to cellular homeostasis and metabolism helping to execute cellular responses to a variety of growth factors and hormones.

Pathways

PI 3 Kinase p85 alpha integrates into two principal signaling networks: the PI3K-Akt and mTOR pathways. These pathways are instrumental for regulating anabolic processes and cell survival. The p85 form of PI3K interacts with additional proteins such as Akt and mTOR coordinating cellular energy balance promoting protein synthesis and inhibiting apoptosis. Through these interactions the protein ensures that cells respond accurately to nutritional and stress signals.

Associated diseases and disorders

Alterations in PI 3 Kinase p85 alpha function have been linked to cancer and type 2 diabetes. Aberrant signaling involving PI3K-Akt-mTOR pathways due to mutations in PI3KR1 can contribute to tumor progression and chemoresistance. Moreover dysfunction of this pathway affects insulin signaling connecting p85 alpha to diabetic pathologies. Proteins such as IRS-1 (insulin receptor substrate 1) act alongside p85 alpha in mediating these disease-associated processes emphasizing its importance in maintaining physiological balance.

Product promise

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12 product images

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Lane 1: Wild type HAP1 whole cell lysate (20 μg)
Lane 2: PIK3R1 knockout HAP1 whole cell lysate (20 μg)
Lane 3: Jurkat whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab191606 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
ab191606 was shown to specifically react with PIK3R1 when PIK3R1 knockout samples were used. Wild-type and PIK3R1 knockout samples were subjected to SDS-PAGE. ab191606 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Predicted band size: 83 kDa
Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PI3K p85 with ab191606 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor^®594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab191606 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Lanes 1- 2: Merged signal (red and green). Green - ab191606 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab191606 was shown to react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell line ab265116 (knockout cell lysate Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate ab257029) was used. Wild-type HeLa and PIK3R1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab191606 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: PIK3R1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell line (Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell line ab265116)
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 90 kDa
Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Blocking/Dilution buffer: 5% NFDM/TBST.
Human PI3K p85 alpha full length recombinant protein contain aa1-724 with a His-Tag® (Cat#Recombinant Human PI 3 Kinase p85 alpha protein ab84769). Human PI3K p85 beta full length recombinant protein contain aa1-728 with a His-Tag® (Cat#Recombinant Human PI 3 Kinase p85 beta protein ab125568).
All lanes: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) at 1/20000 dilution
Lane 1: Human PI3K p85 alpha full length recombinant protein at 0.01 µg
Lane 2: Human PI3K p85 beta full length recombinant protein at 0.01 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 41 kDa, 83 kDa
Observed band size: 55-60 kDa
Exposure time: 5s
Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 100% Methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PI3K p85 with ab191606 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor^®594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab191606 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution.
Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what have been described in the literatures (PMID: 8921377, 12649157).
All lanes: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) at 1/1000 dilution
Lane 1: Human fetal liver lysate at 10 µg
Lane 2: Human fetal heart lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 83 kDa
Observed band size: 46 kDa, 85 kDa
Exposure time: 3min
Flow Cytometry (Intracellular) - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PI3K p85 with ab191606 at 1/150 dilution (red) compared with a Rabbit IgG,monoclonal -Isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluorr^® 488) at 1/500 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
PI3K p85 was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab191606 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab191606 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: MCF7 whole cell lysate, 10μg (Input).
Lane 2: ab191606 IP in MCF7 whole cell lysate.
Lane 3: Rabbit IgG, monoclonal - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab191606 in MCF7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
All lanes: Immunoprecipitation - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Predicted band size: 83 kDa
Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 10 seconds; Lane 3 and 4: 3 seconds.
The molecular weight observed is consistent with what have been described in the literatures (PMID: 8921377, 12649157).
All lanes: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) at 1/1000 dilution
Lane 1: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 2: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4: Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 83 kDa
Observed band size: 46 kDa, 85 kDa
Flow Cytometry (Intracellular) - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Overlay histogram showing HepG2 cells fixed in 4% PFA and stained with ab191606 at a dilution of 1/80 (red line). The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-8: 3 minutes; Lane 9-12: 10 seconds.
The molecular weight observed is consistent with what have been described in the literatures (PMID: 8921377, 12649157).
All lanes: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) at 1/1000 dilution
Lane 1: Mouse brain lysate at 10 µg
Lane 2: Mouse heart lysate at 10 µg
Lane 3: Mouse kidney lysate at 10 µg
Lane 4: Mouse spleen lysate at 10 µg
Lane 5: Rat brain lysate at 10 µg
Lane 6: Rat heart lysate at 10 µg
Lane 7: Rat kidney lysate at 10 µg
Lane 8: Rat spleen lysate at 10 µg
Lane 9: C6 (Rat glial tumor cell line) whole cell lysate at 10 µg
Lane 10: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 11: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 12: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 83 kDa
Observed band size: 46 kDa, 85 kDa
Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606)
PI 3 Kinase p85 alpha Western blot staining of Jurkat (human T cell leukemia T lymphocyte) whole cell lysate using rabbit Anti-PI 3 Kinase p85 alpha antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
Compared with Anti-PI 3 Kinase p85 alpha antibody [EP380Y] ab40755, ab191606 has higher sensitivity. We recommend Anti-S100A4 antibody [EPR2761(2)] ab124805 as an alternative for testing western
blot.
We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using Anti-S100A4 antibody [EPR14639(2)] ab197896 in western blot.
Lane 1: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] (Anti-PI 3 Kinase p85 alpha antibody [EP380Y] ab40755) at 1/1000 dilution
Lane 2: Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) at 1/1000 dilution
All lanes: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 85 kDa
Exposure time: 40s