Recombinant Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Product name

Anti-AKT1 + AKT2 antibody [EPR18405]
See all AKT1 + AKT2 primary antibodies
Description

Rabbit monoclonal [EPR18405] to AKT1 + AKT2
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human, Recombinant fragment
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: AKT1 and AKT2 recombinant protein; HeLa, HepG2, A549, MCF-7, PC12 and NIH/3T3 whole cell lysate; Human fetal kidney lysate; Mouse brain lysate, Rat brain and heart lysate. IHC-P: Human kidney, Human gastric adenocarcinoma, Mouse liver and Rat kidney tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18405
Isotype

IgG
Research areas

Alternative Versions
- Alexa Fluor® 647 Anti-AKT1 + AKT2 antibody [EPR18405] (ab225347)
- Anti-AKT1 + AKT2 antibody [EPR18405] - BSA and Azide free (ab238944)
Assay kits
- Akt Kinase Activity Assay Kit (ab139436)
- AKT Activity Assay Kit (ab65786)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab188099 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
Flow Cyt (Intra)		1/200.
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/2000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
ICC/IF		1/500.

Notes
Flow Cyt (Intra) 1/200.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
ICC/IF 1/500.

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
- SwissProt: P31749 Human
- SwissProt: P31751 Human
- SwissProt: P31750 Mouse
- SwissProt: Q60823 Mouse
- SwissProt: P47196 Rat
- SwissProt: P47197 Rat
see all
Alternative names
- AKT 1 antibody
- AKT 2 antibody
- AKT antibody
- AKT1 antibody
- AKT1 kinase antibody
- AKT1m antibody
- AKT2 antibody
- AKT2 kinase antibody
- C AKT antibody
- CWS6 antibody
- HIHGHH antibody
- MGC99656 antibody
- Murine thymoma viral (v akt) homolog 2 antibody
- Oncogene AKT1 antibody
- PKB alpha antibody
- PKB antibody
- PKB beta antibody
- PKBB antibody
- PKBBETA antibody
- PRKBA antibody
- PRKBB antibody
- Protein kinase Akt 2 antibody
- Protein Kinase B Alpha antibody
- Protein kinase B antibody
- Protein kinase B beta antibody
- Proto oncogene c Akt antibody
- Proto-oncogene c-Akt antibody
- RAC antibody
- RAC alpha antibody
- RAC alpha serine/threonine protein kinase antibody
- RAC BETA antibody
- RAC beta serine threonine protein kinase antibody
- RAC PK alpha antibody
- RAC PK beta antibody
- Rac protein kinase alpha antibody
- Rac protein kinase beta antibody
- RAC-ALPHA antibody
- RACbeta antibody
- v akt murine thymoma viral oncogene homolog 1 antibody
- v akt murine thymoma viral oncogene homolog 2 antibody
- V-AKT murine thymoma viral oncogene homolog 1 antibody
- V-AKT murine thymoma viral oncogene homolog 2 antibody
see all

Western blot - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/20000 dilution

Lane 1 : AKT1 recombinant protein fragment (His-Tag®): aa250-481
Lane 2 : AKT2 recombinant protein fragment (His-Tag®): aa282-481
Lane 3 : AKT3 recombinant protein fragment (His-Tag®): aa351-479

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 56 kDa

Exposure time: 10 seconds

Blocking/dilution buffer: 5% NFDM/TBST.

All three Human AKT recombinant protein fragments containing an N-terminal His-Tag® were made in house.
Western blot - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : MCF-7 (Human breast adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 5 seconds

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution

Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 2 : A549 (Human lung carcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 15 seconds

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 3 minutes

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : Rat heart lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 5 : NIH/3T3 (mouse embryo fibroblast) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 15 seconds

Blocking/dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling AKT1 + AKT2 with ab188099 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing cytoplasmic and weakly nuclear staining on HeLa cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab188099 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling AKT1 + AKT2 with ab188099 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Confocal image showing cytoplasmic and nuclear staining on NIH/3T3 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab188099 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus and cytoplasm staining on normal Human kidney is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus and cytoplasm staining on tumor cells of gastric adenocarcinoma is observed.

Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus and cytoplasm staining on hepatocytes of mouse liver is observed.

Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus and cytoplasm staining on rat kidney is observed.

Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling AKT1/2 with ab188099 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730,black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab188099? Please let us know so that we can cite the reference in this datasheet.

ab188099 has been referenced in 4 publications.

Duan F et al. Vitamin K2 promotes PI3K/AKT/HIF-1a-mediated glycolysis that leads to AMPK-dependent autophagic cell death in bladder cancer cells. Sci Rep 10:7714 (2020). PubMed: 32382009
Xie X et al. Fe3O4-solamargine induces apoptosis and inhibits metastasis of pancreatic cancer cells. Int J Oncol 54:905-915 (2019). PubMed: 30483763
Zhou S et al. ADAMTS13 protects mice against renal ischemia-reperfusion injury by reducing inflammation and improving endothelial function. Am J Physiol Renal Physiol 316:F134-F145 (2019). PubMed: 30461292
Liu J et al. Casticin inhibits nasopharyngeal carcinoma growth by targeting phosphoinositide 3-kinase. Cancer Cell Int 19:348 (2019). PubMed: 31889900

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Assay kits

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Relevance

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (4)

Customer reviews and Q&As