Recombinant Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)
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 - BSA and Azide free
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free
See all PI 3 Kinase catalytic subunit alpha/PIK3CA primary antibodies -
Description
Rabbit monoclonal [EP383Y] to PI 3 Kinase catalytic subunit alpha/PIK3CA - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), WB, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
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).
-
General notes
ab185927 is the carrier-free version of ab40776.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP383Y -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Conjugation kits
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab185927 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 |
---|---|---|
ICC/IF |
Use at an assay dependent concentration.
|
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376-Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
|
WB |
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa).
|
|
IP |
Use at an assay dependent concentration.
|
Notes |
---|
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376-Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa). |
IP
Use at an assay dependent concentration. |
Target
-
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
-
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
-
This WB data was generated using the same anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody clone, EP383Y, in a different buffer formulation (cat# ab40776).
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 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)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40776).
-
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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40776).
-
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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40776).
-
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA with unpurified ab40776 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40776).
-
This ICC/IF data was generated using the same anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody clone, EP383Y, in a different buffer formulation (cat# ab40776).
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).
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
-
Datasheet download
Certificate of Compliance
References (0)
ab185927 has not yet been referenced specifically in any publications.