Recombinant Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP383Y] to PI 3 Kinase catalytic subunit alpha/PIK3CA
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y]
See all PI 3 Kinase catalytic subunit alpha/PIK3CA primary antibodies -
Description
Rabbit monoclonal [EP383Y] to PI 3 Kinase catalytic subunit alpha/PIK3CA -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human PI 3 Kinase catalytic subunit alpha/PIK3CA aa 1000 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P42336 -
Positive control
- WB: Jurkat, MCF-7, Raw264.7 and NIH/3T3 cell lysates. ICC/IF: HeLa and Jurkat cells. IP: Jurkat whole cell lysate (ab7899).
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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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP383Y -
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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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40776 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 110 kDa (predicted molecular weight: 110 kDa).
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ICC/IF |
1/100 - 1/250.
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IP |
1/20 - 1/30.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa). |
ICC/IF
1/100 - 1/250. |
IP
1/20 - 1/30. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Phosphorylates PtdIns, PtdIns4P and PtdIns(4,5)P2 with a preference for PtdIns(4,5)P2. -
Involvement in disease
Defects in PIK3CA are associated with colorectal cancer (CRC) [MIM:114500].
Defects in PIK3CA are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
Defects in PIK3CA are a cause of susceptibility to ovarian cancer (OC) [MIM:167000]. Ovarian cancer common malignancy originating from ovarian tissue. Although many histologic types of ovarian neoplasms have been described, epithelial ovarian carcinoma is the most common form. Ovarian cancers are often asymptomatic and the recognized signs and symptoms, even of late-stage disease, are vague. Consequently, most patients are diagnosed with advanced disease.
Defects in PIK3CA may underlie hepatocellular carcinoma (HCC) [MIM:114550].
Defects in PIK3CA are a cause of keratosis seborrheic (KERSEB) [MIM:182000]. A common benign skin tumor. Seborrheic keratoses usually begin with the appearance of one or more sharply defined, light brown, flat macules. The lesions may be sparse or numerous. As they initially grow, they develop a velvety to finely verrucous surface, followed by an uneven warty surface with multiple plugged follicles and a dull or lackluster appearance. -
Sequence similarities
Belongs to the PI3/PI4-kinase family.
Contains 1 C2 domain.
Contains 1 PI3K/PI4K domain. - Information by UniProt
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Database links
- Entrez Gene: 5290 Human
- Entrez Gene: 18706 Mouse
- Entrez Gene: 170911 Rat
- Omim: 171834 Human
- SwissProt: P42336 Human
- SwissProt: P42337 Mouse
- Unigene: 553498 Human
- Unigene: 85701 Human
see all -
Alternative names
- 5-bisphosphate 3-kinase 110 kDa catalytic subunit alpha antibody
- 5-bisphosphate 3-kinase catalytic subunit alpha isoform antibody
- caPI3K antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PI 3 Kinase catalytic subunit alpha/PIK3CA knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab40776 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab40776 was shown to recognize PI 3 Kinase catalytic subunit alpha/PIK3CA when PI 3 Kinase catalytic subunit alpha/PIK3CA knockout samples were used, along with additional cross-reactive bands. Wild-type and PI 3 Kinase catalytic subunit alpha/PIK3CA knockout samples were subjected to SDS-PAGE. ab40776 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 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 hour at room temperature before imaging. -
ab40776 staining PI 3 Kinase catalytic subunit alpha/PIK3CA in the human cell line Jurkat (human acute T cell leukemia) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776) at 1/1000 dilution
Lane 1 : Jurkat Whole Cell Lysate
Lane 2 : Mouse Brain Tissue Lysate
Lane 3 : Rat Brain Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 110 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab40776 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab40776 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776) at 1/5000 dilution (purified)
Lane 1 : Jurkat whole cell lysate
Lane 2 : MCF-7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 110 kDa
Observed band size: 110 kDa
Blocking and dilution buffer: 5% NFDM/TBST -
All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776) at 1/5000 dilution (purified)
Lane 1 : Raw264.7 whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 110 kDa
Observed band size: 110 kDa
Blocking and dilution buffer: 5% NFDM/TBST -
Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA with purified ab40776 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
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ab40776 (purified) at a dilution of 1/20 immunoprecipitating PI 3 Kinase catalytic subunit alpha/PIK3CA in Jurkat whole cell lysate.
Lane 1 (input): Jurkat whole cell lysate (10µg)
Lane 2 (+): ab40776 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40776 in Jurkat whole cell lysate.
For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776) at 1/5000 dilution (unpurified) + Jurkat cell lysate at 10 µg
Predicted band size: 110 kDa
Observed band size: 110 kDa -
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA with unpurified ab40776 at a dilution of 1/100.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (65)
ab40776 has been referenced in 65 publications.
- Luobu Z et al. CircSCAPER contributes to IL-1β-induced osteoarthritis in vitro via miR-140-3p/EZH2 axis. Bone Joint Res 11:61-72 (2022). PubMed: 35103493
- Wang Q et al. Bacillus subtilis Produces Amino Acids to Stimulate Protein Synthesis in Ruminal Tissue Explants via the Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Beta-Serine/Threonine Kinase-Mammalian Target of Rapamycin Complex 1 Pathway. Front Vet Sci 9:852321 (2022). PubMed: 35832333
- Qiu T et al. Astragaloside IV Inhibits the Proliferation of Human Uterine Leiomyomas by Targeting IDO1. Cancers (Basel) 14:N/A (2022). PubMed: 36139584
- Tang D et al. Silencing LMNB1 Contributes to the Suppression of Lung Adenocarcinoma Development. Cancer Manag Res 13:2633-2642 (2021). PubMed: 33776481
- Feng Z & Yu CH PI(3,4)P2-mediated membrane tubulation promotes integrin trafficking and invasive cell migration. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 33947811