STAT4 Recombinant Rabbit Monoclonal Antibody [JJ08-85]
cat.: ET1701-42
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: JJ08-85
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: 86 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human STAT4 aa 700-748.
Positive control: SiHa cell lysate, A549 cell lysate, HeLa cell lysate, MCF7 cell lysate, SW480 cell lysate, HeLa, human colon carcinoma tissue, human tonsils tissue, mouse testis tissue, rat testis tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP

1:1,000
1:100-1:500
1:100-1:500
1:50-1:1,000
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q14765 Human | P42228 Mouse
Entrez Gene: 367264 Rat
Alternative names: Signal transducer and activator of transcription 4 SLEB11 STAT4 STAT4_HUMAN
Images
ET1701-42_1.jpg Fig1: Western blot analysis of STAT4 on different lysates with Rabbit anti-STAT4 antibody (ET1701-42) at 1/1,000 dilution.

Lane 1: SiHa cell lysate
Lane 2: A549 cell lysate
Lane 3: HeLa cell lysate
Lane 4: MCF7 cell lysate
Lane 5: SW480 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 86 kDa
Observed band size: 75 kDa

Exposure time: 6 minutes;

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 (ET1701-42) 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.
ET1701-42_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling STAT4 with Rabbit anti-STAT4 antibody (ET1701-42) 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-STAT4 antibody (ET1701-42) 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.
ET1701-42_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-STAT4 antibody (ET1701-42) at 1/400 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 (ET1701-42) at 1/400 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.
ET1701-42_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsils tissue with Rabbit anti-STAT4 antibody (ET1701-42) at 1/400 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 (ET1701-42) at 1/400 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.
ET1701-42_5.jpg Fig5: Flow cytometric analysis of HeLa cells labeling STAT4.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-42, 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).
ET1701-42_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-STAT4 antibody (ET1701-42) at 1/1,000 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 (ET1701-42) 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.
ET1701-42_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-STAT4 antibody (ET1701-42) at 1/1,000 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 (ET1701-42) 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.
ET1701-42_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse liver tissue (negative) with Rabbit anti-STAT4 antibody (ET1701-42) at 1/1,000 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 (ET1701-42) 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.
ET1701-42_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat liver tissue (negative) with Rabbit anti-STAT4 antibody (ET1701-42) at 1/1,000 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 (ET1701-42) 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”.