Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6402] to PRAS40 - BSA and Azide free
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-PRAS40 antibody [EPR6402] - BSA and Azide free
See all PRAS40 primary antibodies
DescriptionRabbit monoclonal [EPR6402] to PRAS40 - BSA and Azide free
Tested applicationsSuitable for: WBmore details
Unsuitable for: Flow Cyt,ICC or IHC-P
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human PRAS40 aa 150-250 (internal sequence). The exact sequence is proprietary.
- WB: HAP1, HeLa, and MCF-7 whole cell lysates.
Ab248564 is the carrier-free version of ab133584. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab248564 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
Dissociation constant (KD)KD = 1.43 x 10 -11 M Learn more about KD
Storage bufferConstituent: PBS
Concentration information loading...
Our Abpromise guarantee covers the use of ab248564 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Detects a band of approximately 40 kDa (predicted molecular weight: 27 kDa).|
FunctionSubunit of mTORC1, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves a AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-389', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Within mTORC1, AKT1S1 negatively regulates mTOR activity in a manner that is dependent on its phosphorylation state and binding to 14-3-3 proteins. Inhibits RHEB-GTP-dependent mTORC1 activation. Substrate for AKT1 phosphorylation, but can also be activated by AKT1-independent mechanisms. May also play a role in nerve growth factor-mediated neuroprotection.
Tissue specificityWidely expressed with highest levels of expression in liver and heart. Expressed at higher levels in cancer cell lines (e.g. A549 and HeLa) than in normal cell lines (e.g. HEK293).
modificationsPhosphorylated by AKT1. Phosphorylation relieves inhibitory function on mTORC1.
Cellular localizationCytoplasm > cytosol. Found in the cytosolic fraction of the brain.
- Information by UniProt
- 40 kDa proline rich AKT substrate antibody
- 40 kDa proline-rich AKT substrate antibody
- AKT1 S1 antibody
All lanes : Anti-PRAS40 antibody [EPR6402] (ab133584) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : AKT1S1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF-7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 27 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133584).
ab133584 was shown to specifically react with PRAS40 in wild-type HAP1 cells as signal was lost in AKT1S1 (PRAS40) knockout cells. Wild-type and AKT1S1 (PRAS40) knockout samples were subjected to SDS-PAGE. Ab133584 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab248564 has not yet been referenced specifically in any publications.