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Anti-pan-AKT antibody (ab18785)

  • Datasheet
  • SDS
Reviews (1)Q&A (1)References (36)

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Western blot - Anti-pan-AKT antibody (ab18785)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan-AKT antibody (ab18785)
  • Immunocytochemistry/ Immunofluorescence - Anti-pan-AKT antibody (ab18785)
  • Western blot - Anti-pan-AKT antibody (ab18785)
  • Flow Cytometry - Anti-pan-AKT antibody (ab18785)

Key features and details

  • Rabbit polyclonal to pan-AKT
  • Suitable for: Flow Cyt, ICC/IF, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-pan-AKT antibody
    See all pan-AKT primary antibodies
  • Description

    Rabbit polyclonal to pan-AKT
  • Host species

    Rabbit
  • Specificity

    ab18785 detects Akt1, Akt2 and Akt3.
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to residues surrounding aa 80 of Akt (N terminal)(Human) (Peptide available as ab53312.)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neural Signal Transduction
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • PKB / AKT
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Kinases/Phosphatases
    • Other
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Apoptosis
    • Nuclear
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Serine/threonine kinases
    • AKT
    • Cardiovascular
    • Atherosclerosis
    • Apoptosis
    • Cardiovascular
    • Heart
    • Cardiac metabolism
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab18785 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
ICC/IF
WB (1)
Notes
  • Application notes
    IHC-Fr: Use at a concentration of 10 - 40 µg/ml.
    IP: Use at a concentration of 10 - 40 µg/ml.
    WB: Use at a concentration of 1 - 2 µg/ml. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function

      Plays a role as a 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). General protein kinase capable of phosphorylating several known proteins. Phosphorylates TBC1D4. 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 I (IGF-I). Plays a role in glucose transport by mediating insulin-induced translocation of the GLUT4 glucose transporter to the cell surface. Mediates the antiapoptotic effects of IGF-I. Mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. Promotes glycogen synthesis by mediating the insulin-induced activation of glycogen synthase. The activated form can suppress FoxO gene transcription and promote cell cycle progression. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly.
    • Tissue specificity

      Expressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages.
    • Involvement in disease

      Defects in AKT1 are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
      Defects in AKT1 are associated with colorectal cancer (CRC) [MIM:114500].
      Defects in AKT1 are associated with susceptibility to ovarian cancer [MIM:604370]; also called susceptibility to familial breast-ovarian cancer type 1 (BROVCA1).
    • 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.
    • Domain

      Binding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane. The PH domain mediates interaction with TNK2 and Tyr-176 is also essential for this interaction.
      The AGC-kinase C-terminal mediates interaction with THEM4.
    • Post-translational
      modifications

      Phosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Activated TNK2 phosphorylates it on Tyr-176 resulting in its binding to the anionic plasma membrane phospholipid PA. This phosphorylated form localizes to the cell membrane, where it is targeted by PDPK1 and PDPK2 for further phosphorylations on Thr-308 and Ser-473 leading to its activation. Ser-473 phosphorylation by mTORC2 favors Thr-308 phosphorylation by PDPK1. Ser-473 phosphorylation is enhanced by interaction with AGAP2 isoform 2 (PIKE-A). Ser-473 phosphorylation is enhanced in focal cortical dysplasias with Taylor-type balloon cells.
      Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT1 ubiquitination is critical for phosphorylation and activation. When ubiquitinated, it translocates to the plasma membrane, where it becomes phosphorylated. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.
    • Cellular localization

      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.
    • Target information above from: UniProt accession P31749 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 207 Human
      • Entrez Gene: 11651 Mouse
      • Omim: 164730 Human
      • SwissProt: P31749 Human
      • SwissProt: P31750 Mouse
      • Unigene: 525622 Human
      • Unigene: 6645 Mouse
      • Alternative names

        • AKT1 antibody
        • AKT1_HUMAN antibody
        • AKT2 antibody
        • AKT3 antibody
        • PKB alpha antibody
        • PKB antibody
        • PKB beta antibody
        • PKBalpha antibody
        • PRKBA antibody
        • PRKBB antibody
        • PRKBG antibody
        • Protein kinase B antibody
        • Protein kinase B beta antibody
        • Protein kinase B gamma antibody
        • Proto-oncogene c-Akt antibody
        • RAC alpha antibody
        • RAC antibody
        • RAC gamma antibody
        • RAC PK alpha antibody
        • RAC PK beta antibody
        • RAC PK gamma antibody
        • RAC-alpha serine/threonine-protein kinase antibody
        • RAC-PK-alpha antibody
        • STK2 antibody
        see all

      Images

      • Western blot - Anti-pan-AKT antibody (ab18785)
        Western blot - Anti-pan-AKT antibody (ab18785)
        Anti-pan-AKT antibody (ab18785) + Akt expression in NIH3T3 cell lysate

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

      • Immunocytochemistry/ Immunofluorescence - Anti-pan-AKT antibody (ab18785)
        Immunocytochemistry/ Immunofluorescence - Anti-pan-AKT antibody (ab18785)
        Immunocytochemistry/ Immunofluorescence of HeLa cells staining AKT with ab18785 at 1/10 dilution. Cy3 conjugated Goat anti-rabbit IgG at 1/100 was used a secondary. Counterstained with DAPI. 
         
      • Immunocytochemistry/ Immunofluorescence - Anti-pan-AKT antibody (ab18785)
        Immunocytochemistry/ Immunofluorescence - Anti-pan-AKT antibody (ab18785)

        Immunocytochemistry/ Immunofluorescence of HeLa cells staining AKT with ab18785 at 1/10 dilution. Cy3 conjugated Goat anti-rabbit IgG at 1/100 was used a secondary. Counterstained with DAPI. 

      • Western blot - Anti-pan-AKT antibody (ab18785)
        Western blot - Anti-pan-AKT antibody (ab18785)

        Western blot of HeLa cell lysates staining AKT with ab18785

      • Flow Cytometry - Anti-pan-AKT antibody (ab18785)
        Flow Cytometry - Anti-pan-AKT antibody (ab18785)

        Flow cytometry of HeLa cells.

        Pink: Primary ab18785 at 1/10. Secondary Goat anti-rabbit IgG Cy3.

        Green: Goat anti-rabbit IgG Cy3 only. 

        Black: Unstained HeLa Cells

      Protocols

      • Recommended WB protocol for ab18785

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (36)

      Publishing research using ab18785? Please let us know so that we can cite the reference in this datasheet.

      ab18785 has been referenced in 36 publications.

      • Wang S  et al. miR-21 promotes osteoclastogenesis through activation of PI3K/Akt signaling by targeting Pten in RAW264.7 cells. Mol Med Rep 21:1125-1132 (2020). PubMed: 32016444
      • Liu X  et al. PHLPP Sensitizes Multiple Myeloma Cells to Bortezomib Through Regulating LAMP2. Onco Targets Ther 13:401-411 (2020). PubMed: 32021285
      • Wang S  et al. The Effects of Interleukin-33 (IL-33) on Osteosarcoma Cell Viability, Apoptosis, and Epithelial-Mesenchymal Transition are Mediated Through the PI3K/AKT Pathway. Med Sci Monit 26:e920766 (2020). PubMed: 32312946
      • Qiao LX  et al. Silencing of long non-coding antisense RNA brain-derived neurotrophic factor attenuates hypoxia/ischemia-induced neonatal brain injury. Int J Mol Med 46:653-662 (2020). PubMed: 32626923
      • Zou Y  et al. Downregulation of miRNA-328 promotes the angiogenesis of HUVECs by regulating the PIM1 and AKT/mTOR signaling pathway under high glucose and low serum condition. Mol Med Rep 22:895-905 (2020). PubMed: 32626978
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-2 of 2 Abreviews or Q&A

      Question

      1) Abcam catalogue no:

      ab18785

      2) a) Abcam order number (not required):

      ####

      b) Abcam product lot number:

      923169

      3) Description of the problem (high background, wrong band size, more bands, no band etc):

      No band

      4) Antibody storage conditions (temperature/reconstitution etc):

      -20C (as aliquots long term), 4 C short term

      5) a)Sample (Species):

      human

      b) Type of sample (Cell extract/Nuclear extract/Purified protein/Recombinant protein etc).:

      Cell extract

      6) a) Sample preparation:

      Lysis buffer

      Y

      Protease inhibitors

      Y

      Phosphatase inhibitors

      Y

      Reducing agent

      Y

      Boiling for 5 min?

      Y

      b) Amount of protein loaded:

      Protein loaded ug/lane or cells/lane

      150ug


      ) a) Electrophoresis/Gel conditions:


      Reducing or Non Reducing gel

      reducing

      Percentage of gel

      10%

      Volts applied

      100

      Time applied
      1hr

      b) Transfer or blocking conditions:

      Type of membrane


      nitrocellulose

      Protein transfer verified

      Actin,myc,etc


      Blocking agent and concentration


      5% milk

      Blocking time

      1 hr

      Blocking temperature

      RT

      8) Primary antibody (If more than one was used, describe in additional notes):

      Concentration or dilution

      1:250

      Diluent buffer

      1% BSA in TBST

      Incubation time

      overnight
      Incubation temperature

      4 C

      Washing: Buffer Used

      TBST

      Number of washes

      2


      9) Secondary antibody:

      Species Goat

      Reacts against Rabbit ab

      Concentration or dilution

      1:5000

      Diluent buffer

      2.5% milk in TBST
      Incubation time
      1 hr

      Incubation temperature:

      RT
      Fluorochrome or enzyme conjugate

      HRP CONJUGATED

      Washing: Buffer Used

      TBST

      Number of washes

      3

      10) Detection method (ECL, ECL plus, etc.):


      Pierce west pico soln.

      11) Positive and negative controls used:

      Positive control
      cell line HCT116 irradiated to 6 gy and an un-irradiated HCT116 also

      Negative control

      no

      12) a) How many times you have run this staining?


      3 times

      b) Do you obtain the same results every time?

      yes
      C) What steps have you altered to try and optimize the use of this antibody?

      I have changed the conc of primary Ab from 1:250 and 1:500. Also tried with 75ug and 150 ug. of cell lysate.

      Read More

      Abcam community

      Verified customer

      Asked on Oct 09 2012

      Answer

      Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear the customer has had difficulty obtaining satisfactory results from this antibody.

      The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

      I would like to reassure you that ab18785 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, within 6 months of purchase, we will be pleased to provide a credit note or free of charge replacement.

      Reviewing this case, I would like to offer some suggestions to help optimise the results from ab18785. I would also appreciate if you can confirm some further details:

      1. I notice frmo our records that this item was purchased nearly years ago (Nov 2010). I am sorry this is therefore regrettably no longer covered by the 6 month guarantee, and may have degraded by now. Unfortunately, I think it is too long ago to make an exception in this case. We would encourage all customers to contact us as soon as possible if they encounter any problems.

      2. Please confirm which lysis buffer has been used? I can suggest to try RIPA buffer if this has not already been tried. This shoudl provide a suitable protein preparation.

      3. We recommend not to mix blocking agents in an experiment, this can significantly affect the results. Try using BSA only.

      4. Which cell lines have been tested, other than HCT116?

      5. Was the transfer of protein to the membrane and quality of the sample assessed using a loading control?

      6. Is the current vial of secondary antibody working well with other primary antibodies? How long have has the secondary antibody been stored?

      I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

      Read More

      Abcam Scientific Support

      Answered on Oct 09 2012

      Western blot abreview for Anti-pan-AKT antibody

      Below Average
      Abreviews
      Abreviews
      Application
      Western blot
      Sample
      Mouse Tissue lysate - whole (mouse whole retinal lysate)
      Loading amount
      50 µg
      Specification
      mouse whole retinal lysate
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
      Read More

      Abcam user community

      Verified customer

      Submitted Jan 24 2007

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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