Recombinant Anti-AKT3 + AKT1 antibody [E45] - BSA and Azide free (ab227994)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E45] to AKT3 + AKT1 - BSA and Azide free
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-AKT3 + AKT1 antibody [E45] - BSA and Azide free -
Description
Rabbit monoclonal [E45] to AKT3 + AKT1 - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody detects Akt1 and Akt3.
-
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- Squamous lung carcinoma ICC/IF: SK-BR-3 cells
-
General notes
ab227994 is the carrier-free version of ab32038.
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.
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
E45 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Conjugation kits
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227994 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 |
---|---|---|
WB |
Use at an assay dependent concentration.
|
|
IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
|
ICC/IF |
Use at an assay dependent concentration.
|
Notes |
---|
WB
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
Target
-
Cellular localization
AKT3: Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. AKT1: Cytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus. -
Database links
- Entrez Gene: 100137872 Cow
- Entrez Gene: 280991 Cow
- Entrez Gene: 10000 Human
- Entrez Gene: 207 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 23797 Mouse
- Entrez Gene: 24185 Rat
- Entrez Gene: 29414 Rat
see all
Images
-
Immunocytochemistry/ Immunofluorescence analysis of SK-BR-3 (human breast adenocarcinoma epithelial cell) cells labeling AKT3+AKT1 with purified ab32038 at 1/500 dilution (2.1 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32038). -
This IHC data was generated using the same anti-AKT1/3 antibody clone, E45, in a different buffer formulation (cat# ab32038).
Immunohistochemistry (Paraffin-embedded sections) analysis using ab32038 at a dilution of 1/1000 - 1/1250 and squamous lung carcinoma.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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 (7)
ab227994 has been referenced in 7 publications.
- Liao P et al. Palmitoylated SCP1 is targeted to the plasma membrane and negatively regulates angiogenesis. Elife 6: (2017). WB . PubMed: 28440748
- Chen YC et al. High mobility group box 1-induced epithelial mesenchymal transition in human airway epithelial cells. Sci Rep 6:18815 (2016). WB ; Human . PubMed: 26739898
- Wang Y et al. Intrinsic cellular signaling mechanisms determine the sensitivity of cancer cells to virus-induced apoptosis. Sci Rep 6:37213 (2016). PubMed: 27849011
- Zhang Z et al. Upregulated periostin promotes angiogenesis in keloids through activation of the ERK 1/2 and focal adhesion kinase pathways, as well as the upregulated expression of VEGF and angiopoietin-1. Mol Med Rep 11:857-64 (2015). WB ; Human . PubMed: 25369801
- Verma R & Marchese A The endosomal sorting complex required for transport pathway mediates chemokine receptor CXCR4-promoted lysosomal degradation of the mammalian target of rapamycin antagonist DEPTOR. J Biol Chem 290:6810-24 (2015). PubMed: 25605718
- Feng M et al. Therapeutically targeting glypican-3 via a conformation-specific single-domain antibody in hepatocellular carcinoma. Proc Natl Acad Sci U S A 110:E1083-91 (2013). PubMed: 23471984
- Ulici V et al. The role of Akt1 in terminal stages of endochondral bone formation: angiogenesis and ossification. Bone 45:1133-45 (2009). WB ; Mouse . PubMed: 19679212