PRAS40 Recombinant Rabbit Monoclonal Antibody [JE30-60]
cat.: HA721581
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE30-60
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 27 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human PRAS40 aa 151-200 / 256.
Positive control: HeLa cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, HeLa, human brain tissue, mouse liver tissue, rat liver tissue.
Subcellular location: Cytoplasm, cytosol.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
1:1,000
1:200
1:100
1:500-1:1,000
Uniprot #: SwissProt: Q96B36 Human | Q9D1F4 Mouse
Entrez Gene: 292887 Rat
Alternative names: 40 kDa proline rich AKT substrate 40 kDa proline-rich AKT substrate AKT1 S1 AKT1 substrate 1 (proline rich) AKT1 substrate 1 AKT1S 1 AKT1S1 AKTS1_HUMAN Lobe MGC2865 PRAS 40 PRAS PRAS40 Proline rich akt substrate Proline rich Akt substrate 40 kDa Proline-rich AKT1 substrate 1
Images
HA721581_1.jpg Fig1: Western blot analysis of PRAS40 on different lysates with Rabbit anti-PRAS40 antibody (HA721581) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: PC-12 cell lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 27 kDa
Observed band size: 40 kDa

Exposure time: 1 minute 40 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721581) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
HA721581_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling PRAS40 with Rabbit anti-PRAS40 antibody (HA721581) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PRAS40 antibody (HA721581) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721581_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PRAS40 antibody (HA721581) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721581) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721581_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-PRAS40 antibody (HA721581) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721581) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721581_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-PRAS40 antibody (HA721581) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721581) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721581_6.jpg Fig6: Flow cytometric analysis of HeLa cells labeling PRAS40.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721581, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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