Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) Recombinant Rabbit Monoclonal Antibody [PSH01-38]
cat.: HA721672
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | PSH01-38 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 84 kDa/55 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within human PIK3R1 aa 451-500 / 724. |
| Positive control: | NIH/3T3 treated with 1mM sodium orthovanadate for 30 minutes cell lysate, NIH/3T3 cells treated with 1mM Sodium orthovanadate for 30 minutes, 293T overexpress Src cell lysate, 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate. |
| Subcellular location: | Cell-cell junction, cis-Golgi network, cytoplasm, cytosol, membrane, nucleus. |
| Recommended Dilutions:
WB IF-Cell |
1:1,000 1:100 |
| Uniprot #: | SwissProt: P27986 Human | P26450 Mouse |
| 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 PI3 kinase p85 subunit alpha PI3-kinase regulatory subunit alpha PI3-kinase subunit p85-alpha PI3K PI3K p85 PI3K regulatory subunit alpha Pik3r1 PtdIns 3 kinase p85 alpha PtdIns-3-kinase regulatory subunit alpha PtdIns-3-kinase regulatory subunit p85-alpha |
Images
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Fig1:
Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 treated with 1mM sodium orthovanadate for 30 minutes cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 84 kDa Observed band size: 55/85 kDa Exposure time: 3 minutes 10 seconds; ECL: K1801; 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 (HA721672) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of NIH/3T3 cells treated with or without 1mM Sodium orthovanadate for 30 minutes labeling Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) 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-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) 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. |
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Fig3:
Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody (HA721672) at 1/1,000 dilution. Lane 1: 293T overexpress Src cell lysate Lane 2: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate Lane 3: 293T overexpress Src cell lysate, the membrane treated with λpp for 1 hour Lane 4: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 84 kDa Observed band size: 55 kDa Exposure time: 28 seconds; ECL: K1801; 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 (HA721672) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.