Recombinant Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PI 3 Kinase p85 alpha antibody [EPR5513]
See all PI 3 Kinase p85 alpha primary antibodies -
Description
Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HeLa, Jurkat, Raji, K562, MCF7, C6, Raw 264.7, PC12, NIH3T3 cell lysates
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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 -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 5.50 x 10 -12 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5513 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133595 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 |
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WB |
1/1000 - 1/10000. Detects a band of approximately 85 kDa (predicted molecular weight: 83 kDa).
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Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 85 kDa (predicted molecular weight: 83 kDa). |
Target
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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. -
Tissue specificity
Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level). -
Sequence similarities
Belongs to the PI3K p85 subunit family.
Contains 1 Rho-GAP domain.
Contains 2 SH2 domains.
Contains 1 SH3 domain. -
Domain
The SH3 domain mediates the binding to CBLB, and to HIV-1 Nef. -
Post-translational
modificationsPolyubiquitinated in T-cells by CBLB; which does not promote proteasomal degradation but impairs association with CD28 and CD3Z upon T-cell activation.
Phosphorylated. Dephosphorylated by PTPRJ. - Information by UniProt
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Database links
- Entrez Gene: 5295 Human
- Entrez Gene: 18708 Mouse
- Entrez Gene: 25513 Rat
- Omim: 171833 Human
- SwissProt: P27986 Human
- SwissProt: P26450 Mouse
- SwissProt: Q63787 Rat
- Unigene: 132225 Human
see all -
Alternative names
- GRB1 antibody
- p85 alpha antibody
- p85 antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PI 3 Kinase p85 alpha knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: THP1 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133595 was shown to specifically react with PI 3 Kinase p85 alpha when PI 3 Kinase p85 alpha knockout samples were used. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. ab133595 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PIK3R1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133595 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 ab265116 (knockout 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. ab133595 and 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 (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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All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : Raji cell lysate
Lane 3 : K562 cell lysate
Lane 4 : MCF7 cell lysate
Lane 5 : C6 cell lysate
Lane 6 : Raw 264.7 cell lysate
Lane 7 : PC12 cell lysate
Lane 8 : NIH3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti Rabbit at 1/2000 dilution
Predicted band size: 83 kDa
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab133595 has been referenced in 14 publications.
- Liu G et al. ARHGAP20 Expression Inhibited HCC Progression by Regulating the PI3K-AKT Signaling Pathway. J Hepatocell Carcinoma 8:271-284 (2021). PubMed: 33907697
- Li Y et al. Protective effect of CTRP6 on cerebral ischemia/reperfusion injury by attenuating inflammation, oxidative stress and apoptosis in PC12 cells. Mol Med Rep 22:344-352 (2020). PubMed: 32377750
- Wang W et al. Girdin interaction with vimentin induces EMT and promotes the growth and metastasis of pancreatic ductal adenocarcinoma. Oncol Rep 44:637-649 (2020). PubMed: 32467989
- Liu B et al. Kuijieyuan Decoction Improved Intestinal Barrier Injury of Ulcerative Colitis by Affecting TLR4-Dependent PI3K/AKT/NF-?B Oxidative and Inflammatory Signaling and Gut Microbiota. Front Pharmacol 11:1036 (2020). PubMed: 32848725
- Kang LH et al. Ganoderic acid A protects lens epithelial cells from UVB irradiation and delays lens opacity. Chin J Nat Med 18:934-940 (2020). PubMed: 33357724