WB result of PI3 Kinase p85α Rabbit mAb Primary antibody: PI3 Kinase p85α Rabbit mAb at 1/1000 dilution Lane 1: HepG2 whole cell lysate 20 µg Lane 2: MCF7 whole cell lysate 20 µg Lane 3: Jurkat whole cell lysate 20 µg Lane 4: Raji whole cell lysate 20 µg Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 85 kDa Observed MW: 85, 55, 43 kDa
PI3 Kinase p85α Recombinant Rabbit mAb (S-328-24)
PI3 Kinase p85α Recombinant Rabbit mAb (S-328-24)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | PI3 Kinase p85α |
| Synonyms | Phosphatidylinositol 3-kinase regulatory subunit alpha, PI3-kinase regulatory subunit alpha, PI3K regulatory subunit alpha, PtdIns-3-kinase regulatory subunit alpha, PI3-kinase subunit p85-alpha, PtdIns-3-kinase regulatory subunit p85-alpha, PIK3R1 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus, Membrane |
| Accession | P27986 |
| Clone Number | S-328-24 |
| Antibody Type | Rabbit mAb |
| Application | WB, IHC-P, ICC, ICFCM, IP |
| Reactivity | Hu, Ms, Rt |
| Predicted Reactivity | Or, Bv |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IP | 1:50 | |
| IHC | 1:250 | |
| ICFCM | 1:500 | |
| ICC | 1:500 |
Background
Cell proliferation and many other cell processes can be regulated through a signaling pathway that involves an enzyme called PI3K. This ‘heterodimeric’ enzyme is made up of two protein subunits, one of which is called p85α and inhibits the other subunit of the enzyme (known as p110) to prevent uncontrolled cell proliferation. Activation of the class 1A PI3K is gated, in part, by the p85α regulatory subunit, which contains a Src homology 3 (SH3) domain, two proline-rich (PR) regions (PR1 and PR2) separated by a Rho-GAP/BCR-homology (BH) domain, and two Src homology 2 (SH2) domains (nSH2, cSH2) flanking an inter-SH2 (iSH2) domain [PMID: 26222500].
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Picture
Western Blot
WB result of PI3 Kinase p85α Rabbit mAb Primary antibody: PI3 Kinase p85α Rabbit mAb at 1/1000 dilution Lane 1: NIH/3T3 whole cell lysate 20 µg Lane 2: RAW 264.7 whole cell lysate 20 µg Lane 3: PC-12 whole cell lysate 20 µg Lane 4: mouse brain lysate 20 µg Lane 5: mouse heart lysate 20 µg Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 85 kDa Observed MW: 85, 43 kDa
WB result of PI3 Kinase p85α Rabbit mAb Primary antibody: PI3 Kinase p85α Rabbit mAb at 1/1000 dilution Lane 1: rat brain lysate 20 µg Lane 2: rat heart lysate 20 µg Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 85 kDa Observed MW: 85, 43 kDa
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized Raji (Human Burkitt's lymphoma B lymphocyte) labelling PI3 Kinase p85α antibody at 1/500 (0.1 μg) dilution/ (red) compared with a Rabbit monoclonal IgG (Black) isotype control. Goat Anti-Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
PI3 Kinase p85α Rabbit mAb at 1/50 dilution (1µg) immunoprecipitating PI3 Kinase p85α in 0.4 mg Raji whole cell lysate. Western blot was performed on the immunoprecipitate using PI3 Kinase p85α Rabbit mAb at 1/1000 dilution. Secondary antibody (HRP) for IP was used at 1/400 dilution. Lane 1: Raji whole cell lysate 20 µg (Input) Lane 2: PI3 Kinase p85α Rabbit mAb IP in Raji whole cell lysate Lane 3: Rabbit monoclonal IgG IP in Raji whole cell lysate Predicted MW: 85 kDa Observed MW: 85,55,43 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human placenta. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human testis. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung cancer. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human ovarian cancer. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse kidney. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat stomach. Anti- PI3 Kinase p85α antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in Raji cells. Anti-PI3 Kinase p85α antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).Counterstain with tubulin (Red).
