PI 3 Kinase p85 alpha Recombinant Rabbit Monoclonal Antibody [SU04-07]
cat.: ET1608-70
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: SU04-07
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 84 kDa
Isotype: IgG
Immunogen: Synthetic peptide within C-terminal human PI 3 Kinase p85 alpha.
Positive control: MCF-7 cell lysate, Raji cell lysate, Hela, MCF-7, HepG2, NIH/3T3, human liver carcinoma tissue, human kidney tissue, mouse kidney tissue, mouse heart tissue.
Subcellular location: Cytoplasm, membrane, nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC

1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:50-1:1,000
1:50-1:100
Uniprot #: SwissProt: P27986 Human | P26450 Mouse | Q63787 Rat
Alternative names: GRB1 p85 alpha p85 P85A_HUMAN Phosphatidylinositol 3 kinase associated p 85 alpha Phosphatidylinositol 3 kinase regulatory 1 Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 1 (p85 alpha) Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha Phosphatidylinositol 3-kinase regulatory subunit alpha Phosphoinositide 3 kinase, regulatory subunit 1 (alpha) PI3 kinase p85 subunit alpha PI3-kinase regulatory subunit alpha PI3-kinase subunit p85-alpha PI3K PI3K regulatory subunit alpha Pik3r1 PtdIns 3 kinase p85 alpha PtdIns-3-kinase regulatory subunit alpha PtdIns-3-kinase regulatory subunit p85-alpha
Images
ET1608-70_1.jpg Fig1: Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) at 1/1,000 dilution.

Lane 1: Raji cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate
Lane 4: Mouse brain tissue lysate

Lysates/proteins at 20(cell lysate),40(tissue lysate) µg/Lane.

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 2 minutes;

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 (ET1608-70) 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.
ET1608-70_2.jpg Fig2: ICC staining of PI 3 Kinase p85 alpha in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-70_3.jpg Fig3: ICC staining of PI 3 Kinase p85 alpha in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-70_4.jpg Fig4: ICC staining of PI 3 Kinase p85 alpha in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-70_5.jpg Fig5: ICC staining of PI 3 Kinase p85 alpha in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-70_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-70, 1/50) for 30 minutes 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.
ET1608-70_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-70, 1/50) for 30 minutes 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.
ET1608-70_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-70, 1/50) for 30 minutes 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.
ET1608-70_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-70, 1/50) for 30 minutes 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.
ET1608-70_10.jpg Fig10: Flow cytometric analysis of PI 3 Kinase p85 alpha was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-70, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1608-70_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) 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 (ET1608-70) 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.
ET1608-70_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) at 1/1,000 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 (ET1608-70) at 1/1,000 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.
ET1608-70_13.jpg Fig13: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) at 1/1,000 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 (ET1608-70) at 1/1,000 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.
ET1608-70_14.jpg Fig14: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-PI 3 Kinase p85 alpha antibody (ET1608-70) at 1/1,000 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 (ET1608-70) at 1/1,000 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.
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