Phospho-STAT5 (Y694) Recombinant Rabbit Monoclonal Antibody [SC05-31]
cat.: ET1610-48
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Rat, Mouse |
| Applications: | WB, IHC-P, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | SC05-31 |
| 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: 91 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Tyr694 of human STAT5A. |
| Positive control: | PC-12 treated with 1mM Sodium orthovanadate for 30 minutes cell lysate, PC-12 cells treated with 1mM Sodium orthovanadate for 30 minutes, Hela cells treated with 100ng/mL IFN alpha for 60 minutes, human breast carcinoma tissue. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Cell |
1:1,000-1:2,000 1:50-1:100 1:100 |
| Uniprot #: | SwissProt: P42229 Human | P51692 Human | P42230 Mouse | P42232 Mouse | P52632 Rat | Q62771 Rat |
| Alternative names: | MGF Signal transducer and activator of transcription 5A Signal Transducer and Activator of Transcription 5B STA5A_HUMAN STAT 5A STAT 5B STAT5 STAT5A STAT5B Transcription factor STAT5A Transcription factor STAT5B |
Images
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Fig1:
Western blot analysis of Phospho-STAT5 (Y694) on different lysates with Rabbit anti-Phospho-STAT5 (Y694) antibody (ET1610-48) at 1/1,000 dilution. Lane 1: PC-12 cell lysate Lane 2: PC-12 treated with 1mM Sodium orthovanadate for 30 minutes cell lysate Lane 3: PC-12 treated with 1mM Sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 91 kDa Observed band size: 91 kDa Exposure time: 3 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 (ET1610-48) 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. |
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Fig2:
Immunocytochemistry analysis of PC-12 cells treated with 1mM Sodium orthovanadate for 30 minutes labeling Phospho-STAT5 (Y694) with Rabbit anti-Phospho-STAT5 (Y694) antibody (ET1610-48) 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-Phospho-STAT5 (Y694) antibody (ET1610-48) 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. |
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Fig3:
Western blot analysis of Phospho-STAT5(Y694) on Hela cell lysates. Lane 1: Hela cells, whole cell lysate, 10ug/lane Lane 2/3: Hela cells treated with 100ng/mL IFN alpha for 60 minutes, whole cell lysates, 10ug/lane Lane 4: Hela cells treated with 100ng/mL IFN alpha for 60 minutes, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane All lanes : Anti-Phospho-STAT5(Y694) antibody (ET1610-48) at 1:500 dilution. Anti-STAT5 antibody (ET1612-63) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution. Predicted band size: 91 kDa Observed band size: 91 kDa Blocking and diluting buffer: 5% BSA. Exposure time: 2 minutes 2 seconds |
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Fig4: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-STAT5 (Y694) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1610-48, 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.