PRODUCT CODE: ET1608-70

PI 3 Kinase p85 alpha Recombinant Rabbit Monoclonal Antibody [SU04-07] (ET1608-70)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of PI 3 Kinase p85 alpha on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-70, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: MCF-7 cell lysate<br />
Lane 2: Raji cell lysate
  • Western blot analysis of PI 3 Kinase p85 alpha on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-70, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: MCF-7 cell lysate<br />
Lane 2: Raji cell lysate
  • 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).
  • 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).
  • 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).
  • 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).
  • 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.
  • 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.
  • 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.
  • 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.
  • 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).
Western blot analysis of PI 3 Kinase p85 alpha on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-70, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: MCF-7 cell lysate
Lane 2: Raji cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

PI 3 Kinase p85 alpha Recombinant Rabbit Monoclonal Antibody [SU04-07] (ET1608-70)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Raji, MCF-7, Hela, MCF-7, HepG2, NIH/3T3, mouse kidney tissue, mouse heart tissue, human liver cancer tissue, human kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SU04-07

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

84 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

PI 3 Kinase p85 alpha

SYNONYMS

GRB1 antibody; p85 alpha antibody; p85 antibody; P85A_HUMAN antibody; Phosphatidylinositol 3 kinase associated p 85 alpha antibody; Phosphatidylinositol 3 kinase regulatory 1 antibody; Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 1 (p85 alpha) antibody; Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha antibody; Phosphatidylinositol 3-kinase regulatory subunit alpha antibody; Phosphoinositide 3 kinase, regulatory subunit 1 (alpha) antibody; PI3 kinase p85 subunit alpha antibody; PI3-kinase regulatory subunit alpha antibody; PI3-kinase subunit p85-alpha antibody; PI3K antibody; PI3K regulatory subunit alpha antibody; Pik3r1 antibody; PtdIns 3 kinase p85 alpha antibody; PtdIns-3-kinase regulatory subunit alpha antibody; PtdIns-3-kinase regulatory subunit p85-alpha antibody

SEQUENCE SIMILARITIES

Belongs to the PI3K p85 subunit family.

TISSUE SPECIFICITY

Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level).

POST-TRANSLATIONAL MODIFICATION

Polyubiquitinated in T-cells by CBLB; which does not promote proteasomal degradation but impairs association with CD28 and CD3Z upon T-cell activation.; Phosphorylated. Tyrosine phosphorylated in response to signaling by FGFR1, FGFR2, FGFR3 and FGFR4. Phosphorylated by CSF1R. Phosphorylated by ERBB4. Phosphorylated on tyrosine residues by TEK/TIE2. Dephosphorylated by PTPRJ. Phosphorylated by PIK3CA at Ser-608; phosphorylation is stimulated by insulin and PDGF. The relevance of phosphorylation by PIK3CA is however unclear (By similarity). Phosphorylated in response to KIT and KITLG/SCF. Phosphorylated by FGR.

SUBCELLULAR LOCATION

Cytoplasm, membrane, nucleus.

FUNCTION

Phosphatidylinositol 3-kinase (PI 3-kinase) phosphorylates the 3' OH position of the inositol ring of inositol lipids and is composed of p85 and p110 subunits. PI 3-kinase p85 lacks PI 3-kinase activity and acts as an adapter, coupling p110 to activated protein tyrosine kinase. Two forms of p85 have been described (p85α and p85β), each possessing one SH3 and two SH2 domains. PI 3-kinase p85α, also known as GRB1, phosphatidylinositol 3-kinase regulatory 1 or p85, is a 724 amino acid protein that exists as four alternatively spliced isoforms. Involved in insulin metabolism, defects in the PI 3-kinase p85α gene have been linked to insulin resistance. PI 3-kinase p85α is polyubiquitinated in T-cells by Cbl-b, and has multiple phosphorylated amino acid residues, including a phosphorylated tyrosine residue at position 467.

CITATIONS

  • Cao, Pei et al.

    The age-related changes and differences in energy metabolism and glutamate-glutamine recycling in the d-gal-induced and naturally occurring senescent astrocytes in vitro. | Experimental Gerontology [2019]