Recombinant Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)
![Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-409563-anti-pi-3-kinase-p85-beta-antibody-epr18416-western-blot-hek-293t-lysates.jpg "Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18416] to PI 3 Kinase p85 beta
- Suitable for: IHC-P, WB, ICC/IF, IP
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PI 3 Kinase p85 beta antibody [EPR18416]
See all PI 3 Kinase p85 beta primary antibodies -
Description
Rabbit monoclonal [EPR18416] to PI 3 Kinase p85 beta -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IF, IPmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, Jurkat, HEK293 cell lysates; Human fetal brain and fetal kidney lysates; Rat brain and spleen lysates; PC12 cell lysate. IHC-P: Human colonic adenocarcinoma, Human colon, rat testis tissues. ICC/IF: HeLa and Jurkat cells. IP: Jurkat cell lysate.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18416 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab180967 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| WB |
1/2000. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa).
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| ICC/IF |
1/500.
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| IP |
1/80.
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| Notes |
|---|
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IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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WB
1/2000. Detects a band of approximately 82 kDa (predicted molecular weight: 82 kDa). |
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ICC/IF
1/500. |
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IP
1/80. |
Target
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Function
Regulatory subunit of phosphoinositide-3-kinase (PI3K), a kinase that phosphorylates PtdIns(4,5)P2 (Phosphatidylinositol 4,5-bisphosphate) to generate phosphatidylinositol 3,4,5-trisphosphate (PIP3). PIP3 plays a key role by recruiting PH domain-containing proteins to the membrane, including AKT1 and PDPK1, activating signaling cascades involved in cell growth, survival, proliferation, motility and morphology. Binds to activated (phosphorylated) protein-tyrosine kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Indirectly regulates autophagy (PubMed:23604317). Promotes 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. -
Involvement in disease
Megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome 1 -
Sequence similarities
Belongs to the PI3K p85 subunit family.
Contains 1 Rho-GAP domain.
Contains 2 SH2 domains.
Contains 1 SH3 domain. -
Domain
The SH2 2 domain is required for interaction with FBXL2 and PTPN13. -
Post-translational
modificationsPhosphorylated in response to signaling from activated receptor-type protein kinases (PubMed:19690332, PubMed:20068231). Dephosphorylated by PTPRJ (PubMed:18348712). Dephosphorylated at Tyr-655 by PTPN13. Phosphorylation of Tyr-655 impairs while its dephosphorylation promotes interaction with FBXL2 and SCF(FBXL2)-mediated polyubiquitination (PubMed:23604317).
Ubiquitinated. Polyubiquitination by the SCF(FBXL2) complex probably promotes proteasomal degradation of PIK3R2. - Information by UniProt
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Database links
- Entrez Gene: 5296 Human
- Entrez Gene: 29741 Rat
- Omim: 603157 Human
- SwissProt: O00459 Human
- SwissProt: Q63788 Rat
- Unigene: 371344 Human
- Unigene: 22497 Rat
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Alternative names
- p85 antibody
- p85 beta antibody
- P85B antibody
see all
Images
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Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)All lanes : Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967) at 1/2000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PIK3R2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 82 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab180967 observed at 85 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab180967 was shown to react with PI 3 Kinase p85 beta in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266799 (knockout cell lysate ab257586) was used. Wild-type HEK-293T and PIK3R2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab180967 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246102-anti-pi-3-kinase-p85-beta-antibody-epr18416-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PI 3 Kinase p85 beta using ab180967 at 1/100 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on epithelial cells of Human colon was observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246099-anti-pi-3-kinase-p85-beta-antibody-epr18416-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PI 3 Kinase p85 beta with ab180967 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear and cytoplasmic staining on HeLa cell line.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
1. ab180967 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
![Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-276546-anti-pi-3-kinase-p85-beta-antibody-epr18416-western-blot.jpg)
Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: PIK3R2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab180967 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.ab180967 was shown to specifically react with PIK3R2 (KO) when PIK3R2 (KO) knockout samples were used. Wild-type and PIK3R2 (KO) knockout samples were subjected to SDS-PAGE. Ab180967 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246105-anti-pi-3-kinase-p85-beta-antibody-epr18416-western-blot.jpg)
Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)All lanes : Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967) at 1/2000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 3 : HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 82 kDa
Observed band size: 82 kDa
Exposure time: 30 seconds5% NFDM/TBST: Blocking and diluting buffer.
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![Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246104-anti-pi-3-kinase-p85-beta-antibody-epr18416-western-blot.jpg)
Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)All lanes : Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967) at 1/2000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 82 kDa
Observed band size: 82 kDa
Exposure time: 3 minutes5% NFDM/TBST: Blocking and diluting buffer.
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![Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246103-anti-pi-3-kinase-p85-beta-antibody-epr18416-western-blot.jpg)
Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)All lanes : Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967) at 1/2000 dilution
Lane 1 : Rat brain lysate
Lane 2 : Rat spleen lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 82 kDa
Observed band size: 82 kDa
Exposure time: 30 seconds5% NFDM/TBST: Blocking and diluting buffer.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246101-anti-pi-3-kinase-p85-beta-antibody-epr18416-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)Immunohistochemical analysis of paraffin-embedded Human colonic adenocarcinoma tissue labeling PI 3 Kinase p85 beta using ab180967 at 1/100 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Nucleus and cytoplasm staining on tumor cells of colonic adenocarcinoma was observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246100-anti-pi-3-kinase-p85-beta-antibody-epr18416-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PI 3 Kinase p85 beta using ab180967 at 1/100 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Nucleus and cytoplasm staining on spermatogenic cell of rat testis was observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246098-anti-pi-3-kinase-p85-beta-antibody-epr18416-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PI 3 Kinase p85 beta with ab180967 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear and weakly cytoplasmic staining on Jurkat cell line.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
1. ab180967 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution. -
![Immunoprecipitation - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-246096-anti-pi-3-kinase-p85-beta-antibody-epr18416-immunoprecipitation.jpg)
Immunoprecipitation - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)PI 3 Kinase p85 beta was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab180967 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab180967 at 1/10000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate10 µg (Input).
Lane 2: ab180967 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab180967 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 10 seconds.
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![Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)](https://www.abcam.com/ps/products/180/ab180967/Images/ab180967-13-benefits-of-recombinant-antibodies.png)
Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (18)
ab180967 has been referenced in 18 publications.
- Wang H et al. MiR-126-3p-Enriched Extracellular Vesicles from Hypoxia-Preconditioned VSC 4.1 Neurons Attenuate Ischaemia-Reperfusion-Induced Pain Hypersensitivity by Regulating the PIK3R2-Mediated Pathway. Mol Neurobiol 58:821-834 (2021). PubMed: 33029740
- Zhao HY et al. L-carnitine treatment attenuates renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction. Korean J Intern Med 36:S180-S195 (2021). PubMed: 32942841
- Chen D et al. Suppressive effect of platycodin D on bladder cancer through microRNA-129-5p-mediated PABPC1/PI3K/AKT axis inactivation. Braz J Med Biol Res 54:e10222 (2021). PubMed: 33470388
- Jiang M et al. Hypoxic exposure activates the B cell-specific Moloney murine leukaemia virus integration site 1/PI3K/Akt axis and promotes EMT in leukaemia stem cells. Oncol Lett 21:98 (2021). PubMed: 33376531
- Liu K et al. PHLDA3 inhibition attenuates endoplasmic reticulum stress-induced apoptosis in myocardial hypoxia/reoxygenation injury by activating the PI3K/AKT signaling pathway. Exp Ther Med 21:613 (2021). PubMed: 33936270