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Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) is a rabbit monoclonal antibody that is used to detect AKT1 + AKT2 + AKT3 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat, Xenopus laevis, Xenopus tropicalis samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 310 publications

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Images

Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (AB179463), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (AB179463), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (AB179463), expandable thumbnail
  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (AB179463), expandable thumbnail
  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (AB179463), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Tested
Rat
Expected
Tested
Expected
Expected
Tested
Recombinant fragment
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Xenopus laevis
Expected
Tested
Expected
Expected
Expected
Xenopus tropicalis
Predicted
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species
Human
Dilution info
1/100
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse, Rat, Xenopus laevis
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Xenopus tropicalis
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Recombinant fragment
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/10000
Notes

-

Species
Rat
Dilution info
1/10000
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Xenopus laevis
Dilution info
1/10000
Notes

-

Species
Human
Dilution info
1/10000
Notes

-

Species
Recombinant fragment
Dilution info
1/10000
Notes

-

Predicted
Predicted

Species
Xenopus tropicalis
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

-

Expected
Expected

Species
Mouse, Rat, Xenopus laevis
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Xenopus tropicalis
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Recombinant fragment
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/330
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species
Mouse, Rat, Xenopus laevis
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Xenopus tropicalis
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Recombinant fragment
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/250
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Rat
Dilution info
1/250
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Human
Dilution info
1/250
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species
Xenopus laevis
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Xenopus tropicalis
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Recombinant fragment
Dilution info
-
Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

1 product for Alternative Version

Target data

Function

AKT1 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis (PubMed:11882383, PubMed:15526160, PubMed:15861136, PubMed:21432781, PubMed:21620960, PubMed:31204173). This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates (PubMed:11882383, PubMed:15526160, PubMed:21432781, PubMed:21620960, PubMed:29343641, PubMed:31204173). Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported (PubMed:11882383, PubMed:15526160, PubMed:21432781, PubMed:21620960). AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface (By similarity). Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling (By similarity). Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport (PubMed:11994271). AKT also regulates the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity (By similarity). Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven (By similarity). AKT also regulates cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase) (PubMed:11154276). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis (PubMed:11154276). AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating the mTORC1 signaling pathway, and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1 (PubMed:12150915, PubMed:12172553). Also regulates the mTORC1 signaling pathway by catalyzing phosphorylation of CASTOR1 and DEPDC5 (PubMed:31548394, PubMed:33594058). AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation (By similarity). Part of a positive feedback loop of mTORC2 signaling by mediating phosphorylation of MAPKAP1/SIN1, promoting mTORC2 activation (By similarity). AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization (PubMed:10358075). In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319' (PubMed:10358075). FOXO3 and FOXO4 are phosphorylated on equivalent sites (PubMed:10358075). AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein) (PubMed:9829964). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1 (PubMed:9829964). AKT phosphorylates 'Ser-454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis (By similarity). Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis (By similarity). Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI(3)P-5 activity (By similarity). The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth (By similarity). Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor 1 (IGF1) (PubMed:12176338, PubMed:12964941). AKT mediates the antiapoptotic effects of IGF1 (By similarity). Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly (PubMed:19934221). May be involved in the regulation of the placental development (By similarity). Phosphorylates STK4/MST1 at 'Thr-120' and 'Thr-387' leading to inhibition of its: kinase activity, nuclear translocation, autophosphorylation and ability to phosphorylate FOXO3 (PubMed:17726016). Phosphorylates STK3/MST2 at 'Thr-117' and 'Thr-384' leading to inhibition of its: cleavage, kinase activity, autophosphorylation at Thr-180, binding to RASSF1 and nuclear translocation (PubMed:20086174). Phosphorylates SRPK2 and enhances its kinase activity towards SRSF2 and ACIN1 and promotes its nuclear translocation (PubMed:19592491). Phosphorylates RAF1 at 'Ser-259' and negatively regulates its activity (PubMed:10576742). Phosphorylation of BAD stimulates its pro-apoptotic activity (PubMed:10926925). Phosphorylates KAT6A at 'Thr-369' and this phosphorylation inhibits the interaction of KAT6A with PML and negatively regulates its acetylation activity towards p53/TP53 (PubMed:23431171). Phosphorylates palladin (PALLD), modulating cytoskeletal organization and cell motility (PubMed:20471940). Phosphorylates prohibitin (PHB), playing an important role in cell metabolism and proliferation (PubMed:18507042). Phosphorylates CDKN1A, for which phosphorylation at 'Thr-145' induces its release from CDK2 and cytoplasmic relocalization (PubMed:16982699). These recent findings indicate that the AKT1 isoform has a more specific role in cell motility and proliferation (PubMed:16139227). Phosphorylates CLK2 thereby controlling cell survival to ionizing radiation (PubMed:20682768). Phosphorylates PCK1 at 'Ser-90', reducing the binding affinity of PCK1 to oxaloacetate and changing PCK1 into an atypical protein kinase activity using GTP as donor (PubMed:32322062). Also acts as an activator of TMEM175 potassium channel activity in response to growth factors: forms the lysoK(GF) complex together with TMEM175 and acts by promoting TMEM175 channel activation, independently of its protein kinase activity (PubMed:32228865). Acts as a regulator of mitochondrial calcium uptake by mediating phosphorylation of MICU1 in the mitochondrial intermembrane space, impairing MICU1 maturation (PubMed:30504268). Acts as an inhibitor of tRNA methylation by mediating phosphorylation of the N-terminus of METTL1, thereby inhibiting METTL1 methyltransferase activity (PubMed:15861136). In response to LPAR1 receptor pathway activation, phosphorylates Rabin8/RAB3IP which alters its activity and phosphorylates WDR44 which induces WDR44 binding to Rab11, thereby switching Rab11 vesicular function from preciliary trafficking to endocytic recycling (PubMed:31204173).

Additional Targets

RAC-beta serine/threonine-protein kinase, AKT3

Alternative names

PKB, RAC, AKT1, RAC-alpha serine/threonine-protein kinase, Protein kinase B, Protein kinase B alpha, Proto-oncogene c-Akt, RAC-PK-alpha, PKB alpha

Recommended products

Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) is a rabbit monoclonal antibody that is used to detect AKT1 + AKT2 + AKT3 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat, Xenopus laevis, Xenopus tropicalis samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 310 publications

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR16798
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

What is this antibody validated in?


Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat, Xenopus laevis, Xenopus tropicalis samples.

What is the molecular weight of AKT1 + AKT2 + AKT3?


Anti-AKT1 + AKT2 + AKT3 [EPR16798] (ab179463) specifically detects a band for AKT1 + AKT2 + AKT3 (UniProt: P31749) at a molecular weight of 56kDa.

Trusted by the scientific community


Anti-AKT1 + AKT2 + AKT3 [EPR16798] (ab179463) was first used in a scientific publication in 2014 and has been cited over 310 times in peer-reviewed journals.

Trial sizes available!


Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products


We have a range of other formats of antibody clone [EPR16798] also available for your convenience:
ab179463, PE - PE Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab210454, Carrier free - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] - Low endotoxin, Azide free ab214167, Carrier free - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] - BSA and Azide free ab271930, Alexa Fluor® 647 - Alexa Fluor® 647 Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] ab302553



Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

AKT1 AKT2 and AKT3 also known as Protein Kinase B (PKB) isoforms are serine/threonine-specific protein kinases with critical roles in cellular processes. AKT1 has a molecular weight of about 55.8 kDa AKT2 weighs approximately 56.1 kDa and AKT3 typically has a similar mass. These proteins are expressed in many tissues including brain and heart with AKT1 ubiquitously present AKT2 focused in insulin-responsive tissues and AKT3 mainly in the brain. The molecular weight of AKT plays an important role in their functionality and specificity in tissues.

Biological function summary

AKT proteins regulate cell cycle growing cell survival proliferation and metabolism. They participate as core components of the PI3K/AKT/mTOR signaling pathway forming complexes with other proteins to transmit signals. They bind to phosphoinositide lipids on the cell membrane facilitating their activation and downstream signaling. Through these activities the AKT transporter proteins maintain cellular homeostasis and play a part in stress response.

Pathways

AKT proteins engage in important signaling networks including the PI3K/AKT pathway and mTOR pathway. They work closely with PI3K and mTOR proteins coordinating cellular growth and energy metabolism. In particular the AKT pathway responds to growth factors and insulin influencing glucose uptake and glycolysis regulation through interaction with proteins such as glycogen synthase kinase 3 (GSK3).

Associated diseases and disorders

Dysregulation of AKT signaling can lead to cancer and diabetes. High AKT activation correlates with various cancers by promoting cell survival and growth. In diabetes impaired AKT2 regulation disrupts glucose uptake affecting insulin response. AKT's relationship with mTOR is significant as it often influences tumor growth and progression in cancerous tissues.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    AKT1 + AKT2 + AKT3 Western blot staining using rabbit Anti-AKT1 + AKT2 + AKT3 antibody

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/10000 dilution

    Lane 1: MCF7 (Human breast adenocarcinoma cell line) whole cell lysates at 20 µg

    Lane 2: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg

    Lane 3: Hep G2 (Human liver hepatocellular carcinoma) whole cell lysates at 20 µg

    Lane 4: A549 (Human lung carcinoma) whole cell lysates at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa

    Observed band size: 56 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling AKT1 + AKT2 + AKT3 with ab179463 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on K562 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab179463 at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AKT1 + AKT2 + AKT3 with ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Human renal cortex is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    AKT1 + AKT2 + AKT3 Western blot staining using rabbit Anti-AKT1 + AKT2 + AKT3 antibody

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/1000 dilution

    Lane 1: Human fetal brain lysate at 10 µg

    Lane 2: Human fetal heart lysate at 10 µg

    Lane 3: Human fetal kidney lysate at 10 µg

    Secondary

    All lanes: 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

  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    AKT1 + AKT2 + AKT3 Western blot staining using rabbit Anti-AKT1 + AKT2 + AKT3 antibody

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/1000 dilution

    Lane 1: Mouse brain lysate at 10 µg

    Lane 2: Mouse heart lysate at 10 µg

    Lane 3: Mouse kidney lysate at 10 µg

    Lane 4: Mouse spleen lysate at 10 µg

    Lane 5: Rat brain lysate at 10 µg

    Lane 6: Rat heart lysate at 10 µg

    Lane 7: Rat kidney lysate at 10 µg

    Lane 8: Rat spleen lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa

    Observed band size: 56 kDa

  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    AKT1 + AKT2 + AKT3 Western blot staining of Xenopus muscle lysate using rabbit Anti-AKT1 + AKT2 + AKT3 antibody

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/10000 dilution

    All lanes: Xenopus muscle lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa

    Observed band size: 56 kDa

  • Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    AKT1 + AKT2 + AKT3 Western blot staining using rabbit Anti-AKT1 + AKT2 + AKT3 antibody

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/10000 dilution

    Lane 1: AKT2 recombinant protein (HIS-tag): aa282-481 at 10 µg

    Lane 2: AKT3 recombinant protein (HIS-tag) :aa351-479 at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa

    Observed band size: 18 kDa, 26 kDa

  • Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    AKT1 + AKT2 + AKT3 was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell extract with ab179463 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab179463 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: MCF7 whole cell extract. Lane 2: PBS instead of MCF7 whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    Predicted band size: 56 kDa

    Observed band size: 56 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 with ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Mouse cerebral cortex is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    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 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 with ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Rat cerebral cortex is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)

    Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling AKT1 + AKT2 + AKT3 with ab179463 at 1/330 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

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For this product, it's our understanding that no specific protocols are required. You can:

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