ET1609-30

PI 3 Kinase p85 beta Recombinant Rabbit Monoclonal Antibody [ST04-77]

cat.: ET1609-30

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality:	Monoclonal
Clone number:	ST04-77

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: 82 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human PI 3 Kinase p85 beta aa 136-185 / 728.
Positive control:	Rat heart tissue lysate, PC-12 cell lysate, HeLa cell lysate, NIH/3T3 cell lysates, Hela, SHG-44, human colon tissue, human testis tissue, mouse colon tissue, mouse testis tissue, rat colon tissue, rat testis tissue, HeLa.
Subcellular location:	Cytosol, nucleus
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:1,000-1:2,000 1:50-1:200 1:50-1:200 1:200-1:1,000 1:1,000 1-2μg/sample
Uniprot #:	SwissProt: O00459 Human \| O08908 Mouse \| Q63788 Rat
Alternative names:	p85 p85 beta P85B P85B_HUMAN Phosphatidylinositol 3 kinase Phosphatidylinositol 3 kinase regulatory beta subunit Phosphatidylinositol 3 kinase regulatory subunit beta Phosphatidylinositol 3 kinase regulatory subunit polypeptide 2 Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 2 (p85 beta) Phosphatidylinositol 3-kinase 85 kDa regulatory subunit beta Phosphatidylinositol 3-kinase regulatory subunit beta Phosphoinositide 3 kinase regulatory subunit 2 (beta) Phosphoinositide 3 kinase regulatory subunit 2 Phosphoinositide 3 kinase regulatory subunit polypeptide 2 (p85 beta) Phosphoinositide 3 kinase regulatory subunit polypeptide 2 Phosphoinositide 3 kinase, regulatory subunit 2 (beta) Phosphoinositide 3 kinase, regulatory subunit 2 (p85 beta) PI3 kinase p85 beta subunit PI3 kinase p85 subunit beta PI3-kinase regulatory subunit beta PI3-kinase subunit p85-beta PI3K PI3K regulatory subunit beta PIK3R 2 PIK3R2 PtdIns 3 kinase p......

Images

Fig1: Western blot analysis of PI 3 Kinase p85 beta on different lysates with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

Lane 1: Rat heart tissue lysate
Lane 2: PC-12 cell lysate
Lane 3: HeLa cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 82 kDa
Observed band size: 82 kDa

Exposure time: Lane 1-2: 3 minutes; Lane 3: 1 minute 2 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 (ET1609-30) 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.

Fig2: Western blot analysis of PI 3 Kinase p85 beta on NIH/3T3 cell lysates with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) at 1/1,000 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 82 kDa
Observed band size: 82 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 (ET1609-30) 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 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: ICC staining of PI 3 Kinase p85 beta in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: ICC staining of PI 3 Kinase p85 beta in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig5: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) 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 (ET1609-30) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) 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 (ET1609-30) 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.

	Fig7: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) 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 (ET1609-30) 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) 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 (ET1609-30) 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.
	Fig9: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) 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 (ET1609-30) 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.

	Fig10: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-PI 3 Kinase p85 beta antibody (ET1609-30) 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 (ET1609-30) 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: Flow cytometric analysis of HeLa cells labeling PI 3 Kinase p85 beta. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-30, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig12: PI 3 Kinase p85 beta was immunoprecipitated from 0.2 mg HeLa cell lysate with ET1609-30 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1609-30 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: ET1609-30 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of ET1609-30 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes; ECL: K1801

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