Overview

  • Product name
    Anti-pan-AKT (phospho T308) antibody
    See all pan-AKT primary antibodies
  • Description
    Rabbit polyclonal to pan-AKT (phospho T308)
  • Host species
    Rabbit
  • Specificity
    ab38449 recognises endogenous levels of AKT (pT308), AKT2 (pT309) and AKT3 (pT305) only when phosphorylated at those residues.
  • Tested applications
    Suitable for: WB, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phosphopeptide derived from human AKT1 around the phosphorylation site of Threonine 308 (MKTpFC).

  • Positive control
    • Human lung carcinoma, extracts from 293 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab38449 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
WB 1/500 - 1/1000. Predicted molecular weight: 56 kDa.

Block with BSA.

ELISA 1/10000.
IHC-P Use at an assay dependent concentration.

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.
  • Information by UniProt
  • Database links
  • 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

  • All lanes : Anti-pan-AKT (phospho T308) antibody (ab38449) at 1/500 dilution

    Lane 1 : 293 cell lysate - untreated
    Lane 2 : 293 cell lysate - treated with insulin

    Lysates/proteins at 30 µg per lane.

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

  • Immunohistochemical analysis of AKT (phospho T308) expression in paraffin embedded human lung carcinoma tissue, using ab38449 (1/50). Right-hand panel represents a negative control where ab38449 was pre-incubated with the immunizing (blocking) peptide.

References

This product has been referenced in:
  • Abou-Antoun TJ  et al. Molecular and functional analysis of anchorage independent, treatment-evasive neuroblastoma tumorspheres with enhanced malignant properties: A possible explanation for radio-therapy resistance. PLoS One 13:e0189711 (2018). Read more (PubMed: 29298329) »
  • Yang GS  et al. mTOR is involved in stroke-induced seizures and the anti-seizure effect of mild hypothermia. Mol Med Rep 17:5821-5829 (2018). Read more (PubMed: 29484389) »
See all 73 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Nematostella vectensis Tissue lysate - whole (Whole body)
Gel Running Conditions
Reduced Denaturing (AA 10%)
Loading amount
70 µg
Treatment
200ugr/litr HgCl2
Specification
Whole body
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Vera Brekhman

Verified customer

Submitted Jul 01 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (MRC5 normal lung fibroblasts)
Specification
MRC5 normal lung fibroblasts
Permeabilization
Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Feb 25 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Baboon Tissue lysate - other (Fetal skeletal muscle)
Specification
Fetal skeletal muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Soon-Ok Kim

Verified customer

Submitted Dec 12 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Purified protein (Human primary CD4 T cells)
Specification
Human primary CD4 T cells
Treatment
TCR activated using anti-CD3/CD28 for 3,10 and 30min
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. Marco Craveiro

Verified customer

Submitted Jul 23 2013

Answer


We do have a blocking peptide available for ab38449 however this product is made to order, so it would take approximately 2 months for the order to be processed and shipped.
Please let me know if you would be interested in this product and if so, I will ask our commercial team to add this product to the catalogue.

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Question

To whom it may concern While the picture on the abreview shows this antibody can detected pAkt/T308 (which is the main reason I placed an order for this item- so far, with the 10+ different Ab that I purchased from Abcam based on abreview, this is the first one that I run into problem), I have tried this antibody on 3 different cell lines (CHO-IR, HEK293, R28) in the absence and presence of insulin stimulation without any luck (note- the same samples have been probed for pAkt/Ser473 and I have obtained positive results, hence eliminating the possibility that the insulin stimulation didn't work). After trying the conditions outlined in the Abreview and failing, I have tried the following- blocking (rt, 1 hour)- either 5% milk in TBST(with 0.1% Tween20) or 3% BSA in TBST, also, mixed 5% milk/TBST and 3% BSA.TBST half and half-- didn't work. primary Ab (4 degree, overnight)- have done 1/500, 1/1000 and 1/2000, diluted in 3% BSA/TBST- didn't work I loaded somewhere between 8 to 15 ug of total lysate per lane. The transfer was done using the semi-dry method. For the secondary Ab and ECL combination, I used 1/20,000 (rt for 1 hour, purchased from GE healthcare) for the secondary, which coupled with Pierce ECL substrate-- a combination that I have found to be a good match and have successfully applied to >30 different Abs that I am currently working on, including pAkt/S473 that I mentioned above. I have attached a typical result along with this message. Regardless the condition I tried, I got many non-specific bands (the one between 50 and 37 kDa is always the most intense band!) and can't really see any band that is suppose to be pAkt. Thanks for your input on this matter!

Read More
Answer

Thank you for contacting Abcam.
I am sorry about the issues that you are having with ab38449 in western blotting. Could the band that is in your image you sent at about 55kDa be the correct band? I know it is not in all your plus insulin lanes, so it is probably not, but just wanted to check. However the other non-specific bands do make it difficult to interpret the results.
As the antibody is covered under our Abpromise and it is not giving you the same results that is displayed on the webpages and based on the protocol information you have provided, I do not believe there is anything that I can suggest that would resolve the issue, then I would be happy to replace or refund the cost of the antibody. A suitable replacement antibody would be:
https://www.abcam.com/pan-AKT-phospho-T308-antibody-ab8933.html
If this antibody or any other that is in our catalogue would not meet your needs, I would be happy to refund the antibody. Please let me know how you would like to proceed and if you could also include either the original Abcam order number or PO# that was used to purchase the antibody that would be very helpful.
I look forward to your reply.

Read More

Answer

Thank you for contacting Abcam. We would be very happy if your customer tested ab38449 in ICC/IF in mouse cells. I have placed the discount code and instructions below. If there is anything else I can help you with please let me know. DISCOUNT CODE: **** Expiration date: ******I am very pleased to hear you would like to accept our offer and test ab38449 in ICC/IF in mouse cells. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ICC/IF in mouse cells and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your enquiry. I have contacted the originator of the product but unfortunately, the exact peptide sequence information is proprietary. However, based on the sequence we found for Entamoeba histolytica (swiss prot#Q761W9: Putative uncharacterized protein / Protein kinase AKT), seven amino acids in a row around the phosphorylation site T 308 (TFCGTPE) are the same with the designed peptide, which is around 54% homology. I hope this information will still be useful to you. If there is anything else that I can help you with, please do not hesitate to contact me.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast Canser)
Specification
Breast Canser
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated

Mr. Rudolf Jung

Verified customer

Submitted Mar 09 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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