Estrogen Inducible Protein pS2 Recombinant Rabbit Monoclonal Antibody [JM37-86]
cat.: ET1705-85
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JM37-86
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: 9 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Estrogen Inducible Protein pS2 aa 35-84 / 84.
Positive control: MCF7 cell lysate, MCF7, A549, PC-3M, human stomach tissue, human stomach carcinoma tissue, human breast carcinoma tissue, human pancreas tissue.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:5,000
1:50-1:200
1:50-1:200
1:1,000
1:50-1:100
1:10-1:50
Uniprot #: SwissProt: P04155 Human
Alternative names: BCEI Breast cancer estrogen inducible protein Breast cancer estrogen inducible sequence Breast cancer estrogen-inducible protein D21S21 Gastrointestinal trefoil protein Gastrointestinal trefoil protein pS2 hP1.A HP1A HPS 2 HPS2 pNR 2 PNR-2 pNR2 Polypeptide P1.A Protein pS2 PS 2 pS2 pS2 protein TFF 1 TFF1 TFF1_HUMAN Trefoil factor 1
Images
ET1705-85_1.jpg Fig1: Western blot analysis of Estrogen Inducible Protein pS2 on different lysates with Rabbit anti-Estrogen Inducible Protein pS2 antibody (ET1705-85) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate (15 µg/Lane)
Lane 2: MDA-MB-231 cell lysate (negative) (15 µg/Lane)
Lane 3: Human liver tissue lysate (negative) (20 µg/Lane)
Lane 4: Human kidney tissue lysate (negative) (20 µg/Lane)

Predicted band size: 9 kDa
Observed band size: 9 kDa

Exposure time: 1 minute;

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 (ET1705-85) at 1/5,000 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.
ET1705-85_2.jpg Fig2: Immunocytochemistry analysis of MCF7 cells labeling Estrogen Inducible Protein pS2 with Rabbit anti-Estrogen Inducible Protein pS2 antibody (ET1705-85) 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-Estrogen Inducible Protein pS2 antibody (ET1705-85) 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.
ET1705-85_3.jpg Fig3: ICC staining of Estrogen Inducible Protein pS2 in A549 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 (ET1705-85, 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).
ET1705-85_4.jpg Fig4: ICC staining of Estrogen Inducible Protein pS2 in PC-3M 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 (ET1705-85, 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).
ET1705-85_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-Estrogen Inducible Protein pS2 antibody (ET1705-85) 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 (ET1705-85) 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.
ET1705-85_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Rabbit anti-Estrogen Inducible Protein pS2 antibody (ET1705-85) 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 (ET1705-85) 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.
ET1705-85_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Estrogen Inducible Protein pS2 antibody (ET1705-85) 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 (ET1705-85) 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.
ET1705-85_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Estrogen Inducible Protein pS2 antibody (ET1705-85) 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 (ET1705-85) 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.
ET1705-85_9.jpg Fig9: Flow cytometric analysis of Estrogen Inducible Protein pS2 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1705-85, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1705-85_10.jpg Fig10: Flow cytometric analysis of MCF7 cells labeling Estrogen Inducible Protein pS2.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-85, 1μg/mL) (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).
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