PTEN Recombinant Rabbit Monoclonal Antibody [SJ19-03]
cat.: ET1606-43
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SJ19-03
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: 47 kDa
Isotype: IgG
Immunogen: Recombinant full length protein of human PTEN.
Positive control: MCF7 cell lysate, A431 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, PC-12 cell lysate, NIH/3T3, mouse brain tissue, mouse pancreas tissue, rat brain tissue, rat pancreas tissue.
Subcellular location: Cytoplasm, Nucleus, Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:500-1:5,000
1:100-1:500
1:50
1:200
Uniprot #: SwissProt: P60484 Human | O08586 Mouse
Unigene: 22158 Rat
Alternative names: 10q23del BZS DEC GLM2 MGC11227 MHAM MMAC1 MMAC1 phosphatase and tensin homolog deleted on chromosome 10 Mutated in multiple advanced cancers 1 Phosphatase and tensin homolog Phosphatase and tensin like protein Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN Pten PTEN_HUMAN PTEN1 TEP1
Images
ET1606-43_1.jpg Fig1: Western blot analysis of PTEN on different lysates with Rabbit anti-PTEN antibody (ET1606-43) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: MDA-MB-468 cell lysate (negative)
Lane 3: A431 cell lysate
Lane 4: HeLa cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: C6 cell lysate
Lane 7: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 47 kDa
Observed band size: 55 kDa

Exposure time: 1 minute 10 seconds;

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 (ET1606-43) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were 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.
ET1606-43_2.jpg Fig2: All lanes: Western blot analysis of PTEN with anti-PTEN antibody (ET1606-43) at 1:500 dilution.

Lane 1/2: Wild-type Hela whole cell lysate (10 µg).
Lane 3/4: PTEN knockout Hela whole cell lysate (10 µg).

ET1606-43 was shown to specifically react with PTEN in wild-type Hela cells. No band was observed when PTEN knockout sample was tested. Wild-type and PTEN knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1606-43, 1/500) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) 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.
ET1606-43_3.jpg Fig3: All lanes: Western blot analysis of PTEN with anti-PTEN antibody [SJ19-03] (ET1606-43) at 1:1,000 dilution.

Lane 1: Wild-type mouse lung cancer whole cell lysate (20 µg).
Lane 2: PTEN knockout mouse lung cancer whole cell lysate (20 µg).

ET1606-43 was shown to specifically react with PTEN in wild-type mouse lung cancer cells. No band was observed when PTEN knockout samples were tested. Wild-type and PTEN knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-PTEN antibody (ET1606-43, 1/1,000) and Anti-β-actin antibody (R1207-1, 1/1,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1606-43_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling PTEN with Rabbit anti-PTEN antibody (ET1606-43) 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-PTEN antibody (ET1606-43) 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.
ET1606-43_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-PTEN antibody (ET1606-43) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1606-43) 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.
ET1606-43_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-PTEN antibody (ET1606-43) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1606-43) 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.
ET1606-43_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-PTEN antibody (ET1606-43) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1606-43) 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.
ET1606-43_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-PTEN antibody (ET1606-43) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1606-43) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.