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 1:30,000 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 cancer tissue with Rabbit anti-Phospho-STAT5 (Y694) antibody (ET1610-48) at 1/30,000 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 (ET1610-48) at 1/30,000 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”.