Overview

  • Product name
    Anti-AKT2 antibody [4H7]
    See all AKT2 primary antibodies
  • Description
    Mouse monoclonal [4H7] to AKT2
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, ChIP, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
    Predicted to work with: Non human primates
  • Immunogen

    Recombinant full length protein corresponding to Human AKT2 aa 1-481. (NP_001617) produced in HEK293T cell.
    Sequence:

    MNEVSVIKEGWLHKRGEYIKTWRPRYFLLKSDGSFIGYKERPEAPDQTLP PLNNFSVAECQLMKTERPRPNTFVIRCLQWTTVIERTFHVDSPDEREEWM RAIQMVANSLKQRAPGEDPMDYKCGSPSDSSTTEEMEVAVSKARAKVTMN DFDYLKLLGKGTFGKVILVREKATGRYYAMKILRKEVIIAKDEVAHTVTE SRVLQNTRHPFLTALKYAFQTHDRLCFVMEYANGGELFFHLSRERVFTEE RARFYGAEIVSALEYLHSRDVVYRDIKLENLMLDKDGHIKITDFGLCKEG ISDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRL PFYNQDHERLFELILMEEIRFPRTLSPEAKSLLAGLLKKDPKQRLGGGPS DAKEVMEHRFFLSINWQDVVQKKLLPPFKPQVTSEVDTRYFDDEFTAQSI TITPPDRYDSLGLLELDQRTHFPQFSYSASIRE


    Database link: P31751

  • Positive control
    • MCF7, HeLa, HepG2, A549, 293T, Jurkat, A431, U2OS, COS7, 3T3 L1 and NRK whole celll lysates; AKT2 transfected U2OS cells; Human Medulla Oblongata tissue; Human Esophageal cancer tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab175354 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
ICC/IF 1/10 - 1/100.
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Use 2μg.

IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 55 kDa.

Target

  • Function
    General protein kinase capable of phosphorylating several known proteins.
  • Tissue specificity
    Expressed in all human cell types so far analyzed.
  • Sequence similarities
    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 PH domain.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Phosphorylation on Thr-309 and Ser-474 is required for full activity.
    Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT2 ubiquitination. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.
  • Information by UniProt
  • Database links
  • Alternative names
    • Akt2 antibody
    • AKT2_HUMAN antibody
    • HIHGHH antibody
    • murine thymoma viral (v-akt) homolog-2 antibody
    • PKB antibody
    • PKB beta antibody
    • PKBB antibody
    • PKBBETA antibody
    • PRKBB antibody
    • Protein kinase Akt 2 antibody
    • Protein kinase Akt-2 antibody
    • Protein kinase B beta antibody
    • rac protein kinase beta antibody
    • RAC-BETA antibody
    • RAC-beta serine/threonine-protein kinase antibody
    • RAC-PK-beta antibody
    • v akt murine thymoma viral oncogene homolog 2 antibody
    see all

Images

  • Immunofluorescent analysis of AKT2 (green) showing staining in the cytoplasm and nucleus of C2C12 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an AKT2 monoclonal antibody (ab175354) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunofluorescent analysis of AKT2 (green) showing staining in the cytoplasm and nucleus of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an AKT2 monoclonal antibody (ab175354) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Immunofluorescent analysis of AKT2 (green) showing staining in the cytoplasm and nucleus of MCF-7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an AKT2 monoclonal antibody (ab175354) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

  • Chromatin immunoprecipitation analysis of Akt1 and Akt2 was performed using cross-linked chromatin from 1 x 106 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed with 1.0ul/100ul well volume of an Atk1 monoclonal antibody  and an Akt2 monoclonal antibody (ab175354). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1ul of eluted DNA in 2ul SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 of Egr1. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions); the zigzag line represents an intron; and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars.

  • Immunohistochemical analysis of deparaffinized Human Esophageal cancer tissue labeling AKT2 with ab175354 at 1/200 dilution. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate.

  • All lanes : Anti-AKT2 antibody [4H7] (ab175354) at 1/1000 dilution

    Lane 1 : MCF7 whole cell lysate
    Lane 2 : HeLa whole cell lysate
    Lane 3 : HepG2 whole cell lysate
    Lane 4 : A549 whole cell lysate
    Lane 5 : 293T whole cell lysate
    Lane 6 : Jurkat whole cell lysate
    Lane 7 : A431 whole cell lysate
    Lane 8 : U2OS whole cell lysate
    Lane 9 : COS7 whole cell lysate
    Lane 10 : 3T3 L1 whole cell lysate
    Lane 11 : NRK whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : goat anti-mouse-HRP at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 55 kDa

  • Immunoprecipitation of AKT2 was performed on HeLa cells. The antigen:antibody complex was formed by incubating 750 µg whole cell lysate with 2 µg of ab175354. WB detection used ab175354 at 1/1000 dilution.

  • Immunohistochemical analysis of deparaffinized normal Human Medulla Oblongata tissue labeling AKT2 with ab175354 at 1/200 dilution. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate. 

  • All lanes : Anti-AKT2 antibody [4H7] (ab175354) at 1/1000 dilution

    Lane 1 : Non-transfected U2OS cells
    Lane 2 : AKT2 transfected U2OS cells

    Secondary
    All lanes : goat anti-mouse-HRP at 1/20000 dilution

    Predicted band size: 55 kDa

References

This product has been referenced in:
  • Guan G  et al. Expression of RNA-binding motif 10 is associated with advanced tumor stage and malignant behaviors of lung adenocarcinoma cancer cells. Tumour Biol 39:1010428317691740 (2017). Read more (PubMed: 28347232) »
  • Shen L  et al. C-X-C motif chemokine ligand 8 promotes endothelial cell homing via the Akt-signal transducer and activator of transcription pathway to accelerate healing of ischemic and hypoxic skin ulcers. Exp Ther Med 13:3021-3031 (2017). Read more (PubMed: 28587375) »

See all 4 Publications for this product

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