Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18405] to AKT1 + AKT2
- Suitable for: IHC-P, WB, Flow Cyt, ICC/IF
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Overview
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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 & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF MouseHumanIHC-P MouseRatHumanWB MouseHumanRecombinant fragment -
Immunogen
This information is proprietary to Abcam and/or its suppliers.
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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 Cytometry: HeLa cells.
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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 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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18405 -
Isotype
IgG -
Research areas
Images
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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 secondsBlocking/dilution buffer: 5% NFDM/TBST.
All three Human AKT recombinant protein fragments containing an N-terminal His-Tag® were made in house.
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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 secondsBlocking/dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/dilution buffer: 5% NFDM/TBST.
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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. -
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. -
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.
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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.
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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.
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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.
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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.
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