STAT 5A+B Recombinant Rabbit Monoclonal Antibody [JJ08-78]
cat.: ET1701-45
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JJ08-78
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: 90 kDa
Isotype: IgG
Immunogen: Recombinant protein within mouse Stat5b aa 585-773 / 786.
Positive control: A549 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Mouse testis tissue lysate, Rat ovary tissue lysate, PC-12, HepG2, NIH/3T3, human tonsil tissue, rat lymph node tissue, rat testis tissue, mouse brain tissue, human spleen tissue.
Subcellular location: Nucleus, Cytoplasm.
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
1:50-1:400
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P42229 Human | P51692 Human | P42230 Mouse | P42232 Mouse | P52632 Rat | Q62771 Rat
Alternative names: Mammary gland factor (STAT5A) MGF (STAT5A) Signal transducer and activator of transcription 5A Signal transducer and activator of transcription 5B STA5A_HUMAN Stat 5a STAT 5b STAT5A Transcription factor STAT5B
Images
ET1701-45_1.jpg Fig1: Western blot analysis of STAT 5A+B on different lysates with Rabbit anti-STAT 5A+B antibody (ET1701-45) at 1/1,000 dilution.

Lane 1: A549 cell lysate (10 µg/Lane)
Lane 2: HeLa cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: PC-12 cell lysate (10 µg/Lane)
Lane 5: Mouse brain tissue lysate (20 µg/Lane)
Lane 6: Mouse testis tissue lysate (20 µg/Lane)
Lane 7: Rat ovary tissue lysate (20 µg/Lane)

Predicted band size: 90 kDa
Observed band size: 90 kDa

Exposure time: 4 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 (ET1701-45) 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.
ET1701-45_2.jpg Fig2: Immunocytochemistry analysis of PC-12 cells labeling STAT 5A+B with Rabbit anti-STAT 5A+B antibody (ET1701-45) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-STAT 5A+B antibody (ET1701-45) 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 (HA601187, 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.
ET1701-45_3.jpg Fig3: ICC staining of STAT 5A+B in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-45, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-45_4.jpg Fig4: ICC staining of STAT 5A+B in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-45, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-45_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-STAT 5A+B antibody (ET1701-45) 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 (ET1701-45) 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.
ET1701-45_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat lymph node tissue with Rabbit anti-STAT 5A+B antibody (ET1701-45) 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 (ET1701-45) 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.
ET1701-45_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-STAT 5A+B antibody (ET1701-45) 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 (ET1701-45) 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.
ET1701-45_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-STAT 5A+B antibody (ET1701-45) 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 (ET1701-45) 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.
ET1701-45_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-STAT 5A+B antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-45, 1/50) for 30 minutes 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.
ET1701-45_10.jpg Fig10: Flow cytometric analysis of PC-12 cells labeling STAT 5A+B.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-45, 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).
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