EM1701-62

PI 3 Kinase p85 alpha Mouse Monoclonal Antibody [L3-D6]

cat.: EM1701-62

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, IF-Tissue
Clonality:	Monoclonal
Clone number:	L3-D6

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein G affinity purified.
Molecular weight:	Predicted band size: 84 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within Human PI3-kinase p85 subunit alpha aa 19-219 / 724.
Positive control:	Raji cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HepG2, LOVO, rat brain tissue, human tonsil tissue, human colon cancer tissue, human placenta tissue.
Subcellular location:	Cytosol. Endoplasmic reticulum. Golgi apparatus. Nucleus. Plasma Membrane.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue	1:500-1:2,000 1:50-1:200 1:50-1:1,000 1:200
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

Fig1: Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) at 1/1,000 dilution.

Lane 1: Raji cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 84 kDa
Observed band size: 84 kDa

Exposure time: 2 minutes; 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 (EM1701-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) at 1/1,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si PI 3 Kinase p85 alpha cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 84 kDa
Observed band size: 84 kDa

Exposure time: 3 minutes; ECL: K1802;

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 (EM1701-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: ICC staining PI 3 Kinase p85 alpha (green) in HepG2 cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
	Fig4: ICC staining PI 3 Kinase p85 alpha (green) in LOVO cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
	Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PI 3 Kinase p85 alpha antibody. Counter stained with hematoxylin.
	Fig6: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-PI 3 Kinase p85 alpha antibody. Counter stained with hematoxylin.

	Fig7: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-PI 3 Kinase p85 alpha antibody. Counter stained with hematoxylin.
	Fig8: Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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.
	Fig9: Immunohistochemical analysis of paraffin-embedded human lung tissue with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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.
	Fig10: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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.

Fig11: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Mouse anti-PI 3 Kinase p85 alpha antibody (EM1701-62) 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 (EM1701-62) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.