Recombinant Anti-AKT1 + AKT2 antibody [EPR17062] - BSA and Azide free (ab250634)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17062] to AKT1 + AKT2 - BSA and Azide free
- Suitable for: IP, IHC-P, Flow Cyt (Intra), WB, ICC/IF
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
-
Product name
Anti-AKT1 + AKT2 antibody [EPR17062] - BSA and Azide free
See all AKT1 + AKT2 primary antibodies -
Description
Rabbit monoclonal [EPR17062] to AKT1 + AKT2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-P, Flow Cyt (Intra), WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
General notes
ab250634 is the carrier-free version of ab182729.
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.
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17062 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab250634 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 |
---|---|---|
IP |
Use at an assay dependent concentration.
|
|
IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
|
WB |
Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
|
|
ICC/IF |
Use at an assay dependent concentration.
|
Notes |
---|
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
ICC/IF
Use at an assay dependent concentration. |
Target
-
Relevance
The serine/threonine kinase AKT (protein kinase B or PKB) has a central role in the regulation of several signaling pathways controlling cell proliferation, apoptosis, angiogenesis, and diabetes. In humans, there are three genes in the "AKT family": AKT1, AKT2, and AKT3. AKT1 is catalytically inactive in serum starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet derived growth factor. The activation is rapid and specific. In the developing nervous system AKT is a critical mediator of growth factor induced neuronal survival. Survival factors can suppress apoptosis in a transcription independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. AKT2 is a putative oncogene and is a general protein kinase capable of phophorylating several known proteins. AKT2 is amplified and overexpressed in some human carcinomas. AKT2 acts primarily as a regulator of glucose metabolism. -
Cellular localization
ATK1: Cytoplasm. Nucleus. Cell membrane. Note: Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. -
Database links
- Entrez Gene: 207 Human
- Entrez Gene: 208 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 11652 Mouse
- Entrez Gene: 24185 Rat
- Entrez Gene: 25233 Rat
- Omim: 164730 Human
- Omim: 164731 Human
see all -
Alternative names
- AKT 1 antibody
- AKT 2 antibody
- AKT antibody
see all
Images
-
All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/1000 dilution
Lane 1 : Human AKT1 fragment recombinant protein
Lane 2 : Human AKT2 fragment recombinant protein
Lane 3 : Human AKT3 fragment recombinant protein
Lysates/proteins at 0.02 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Exposure time: 1 secondThis data was developed using ab182729, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Human AKT1 fragment recombinant protein contains aa281-480 with a His-Tag®. Human AKT2 fragment recombinant protein contains aa282-481 with a His-Tag®. Human AKT3 fragment recombinant protein contains aa351-479 with a His-Tag®.
-
All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/5000 dilution
Lane 1 : Human fetal heart lysate
Lane 2 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDa
Exposure time: 30 secondsThis data was developed using ab182729, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/5000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaThis data was developed using ab182729, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1: 8 seconds; Lane 2: 2 seconds.
-
All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/5000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate
Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 6 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 7 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaThis data was developed using ab182729, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: Lanes 1-4: 5 seconds; Lanes 5-7: 3 seconds.
-
This data was developed using ab182729, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Human tonsil is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. -
This data was developed using ab182729, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human bladder cancer tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Human bladder cancer is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. -
This data was developed using ab182729, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Mouse stomach is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. -
This data was developed using ab182729, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Rat cerebrum is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. -
This data was developed using ab182729, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling AKT1 + AKT2 with ab182729 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 ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.
-
This data was developed using ab182729, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling AKT1 + AKT2 with ab182729 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 NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.
-
This data was developed using ab182729, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling AKT1 + AKT2 with ab182729 at 1/600 dilution (red) compared with a 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 Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
-
This data was developed using ab182729, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling AKT1 + AKT2 with ab182729 at 1/600 dilution (red) compared with a 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 Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
-
This data was developed using ab182729, the same antibody clone in a different buffer formulation.AKT1/2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab182729 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab182729 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution. Lane 1: HeLa whole cell lysate, 10µg (Input). Lane 2: ab182729 IP in HeLa whole cell lysate. Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab182729 in HeLa whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 30 seconds.
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)
ab250634 has not yet been referenced specifically in any publications.