Phospho-STAT3 (Y705) Recombinant Rabbit Monoclonal Antibody [SZ43-01]
cat.: ET1603-40
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | SZ43-01 |
| 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: 88 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Tyr705 of human STAT3. |
| Positive control: | HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate, Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate, A431 treated with 100ng/mL EGF for 30 minutes whole cell lysate, human lung tissue, Hela cell lysate, SK-Br-3 cell lysate, human kidney tissue, mouse skeletal muscle tissue. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IHC-P IP |
1:1,000-1:5,000 1:500 1-2μg/sample |
| Uniprot #: | SwissProt: P40763 Human | P42227 Mouse | P52631 Rat |
| Alternative names: | 1110034C02Rik Acute Phase Response Factor Acute-phase response factor ADMIO APRF AW109958 DNA binding protein APRF FLJ20882 HIES MGC16063 Signal transducer and activator of transcription 3 (acute phase response factor) Signal transducer and activator of transcription 3 STAT 3 Stat3 STAT3_HUMAN |
Images
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Fig1:
Western blot analysis of Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate Lane 3: Jurkat whole cell lysate Lane 4: Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate Lane 5: A431 whole cell lysate Lane 6: A431 treated with 100ng/mL EGF for 30 minutes whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 88 kDa Observed band size: 88 kDa Exposure time: 35 seconds; 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 (ET1603-40) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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:
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1603-40) at 1/500 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. |
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Fig3:
Western blot analysis of Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/1,000 dilution. Lane 1: M‑NFS‑60 cell lysate Lane 2: RAW264.7 cell lysate Lane 3: C2C12 cell lysate Lane 4: L6 cell lysate Lane 2: Mouse heart tissue lysate Cell ysates/proteins at 20 µg/Lane. Tissue ysates/proteins at 20 µg/Lane. Predicted band size: 88 kDa Observed band size: 88 kDa Exposure time: 25 seconds; 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 (ET1603-40) 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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Fig4:
Western blot analysis of Phospho-STAT3 (Y705) on SK-Br-3 cell lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution. Lysates/proteins at 10 µg/Lane. Predicted band size: 88 kDa Observed band size: 88 kDa Exposure time: 2 minutes; 8% 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 (ET1603-40) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1603-40) at 1/500 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. |
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Fig6:
Phospho-STAT3 (Y705) was immunoprecipitated from 0.2 mg HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate with ET1603-40 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1603-40 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate (input) Lane 2: Rabbit IgG instead of ET1603-40 in HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate Lane 3: ET1603-40 IP in HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 24 seconds; ECL: K1802 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.