ET1701-45

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:	Jurkat cell lysate,PC-12 cell lysate,HepG2 cell lysate,NIH/3T3 cell lysate,RH-35 cell lysate,HeLa cell lysate,HepG2, NIH/3T3, RH-35, human tonsil tissue, human spleen tissue, mouse brain 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 1:50-1:100 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

	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: Jurkat cell lysate Lane 2: PC-12 cell lysate Lane 3: HepG2 cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: RH-35 cell lysate Lane 6: HeLa cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 90 kDa Observed band size: 90 kDa Exposure time: 2 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 (ET1701-45) 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 1:100,000 dilution was used for 1 hour at room temperature.
	Fig2: 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).
	Fig3: 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).

	Fig4: ICC staining of STAT 5A+B in RH-35 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).
	Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil 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.
	Fig6: 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.
	Fig7: 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.

Fig8: Flow cytometric analysis of STAT 5A+B was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-45, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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