WB
Western blot analysis of Phospho-STAT1 (S727) on different lysates with Rabbit anti-Phospho-STAT1 (S727) antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane), Lane 2: Mouse brain tissue lysate (20 µg/Lane), Lane 3: Rat brain tissue lysate (20 µg/Lane), Lane 4: Mouse brain treated with λpp for 1 hour tissue lysate (20 µg/Lane). Exposure time: 3 minutes 10 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-Phospho-STAT1 (S727) antibody 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 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.IHC
Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-Phospho-STAT1 (S727) antibody 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 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Phospho-STAT1 (S727) antibody 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 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.ICC/IF
Immunocytochemistry analysis of HeLa cells treated with or without λpp labeling Phospho-STAT1 (S727) with Rabbit anti-Phospho-STAT1 (S727) antibody at 1/2,000 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-STAT1 (S727) antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling Phospho-STAT1 (S727). Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).WB
(B) DEF cells were transfected with a lentiviral vector expressing duck RIG-I and a lentivirus encoding ZsGreen for 24 h, and then infected with DEV at an MOI of 1 or mock-infected for 48 h, Expression levels of STAT1 and pSTAT1 in the cell lysates were analyzed by western blotting, with β-actin protein as the control.| Product Name | Phospho-STAT1 (S727) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Membrane receptor signaling by various ligands, including interferons and growth hormones such as EGF, induces activation of Jak kinases which then leads to tyrosine phosphorylation of the various Stat transcription factors. Stat1 and Stat2 are induced by IFN-a and form a heterodimer which is part of the ISGF3 transcription factor complex. Although early reports indicate Stat3 activation by EGF and IL-6, it has been shown that Stat3β appears to be activated by both while Stat3α is activated by EGF, but not by IL-6. Highest expression of Stat4 is seen in testis and myeloid cells. IL-12 has been identified as an activator of Stat4. Stat5 has been shown to be activated by prolactin and by IL-3. Stat6 is involved in IL-4 activated signaling pathways. |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser727 of Human STAT1 aa 701-750 / 750. |
| Modification | p-S727 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:2000-1:5000 IHC1:200-1:1000ICC/IF:1:2000 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Signal transducer and activator of transcription 1 91kD antibody
CANDF7 antibody
DKFZp686B04100 antibody
IMD31A antibody
IMD31B antibody
IMD31C antibody
ISGF 3 antibody
ISGF-3 antibody
OTTHUMP00000163552 antibody
OTTHUMP00000165046 antibody
OTTHUMP00000165047 antibody
OTTHUMP00000205845 antibody
Signal transducer and activator of transcription 1 91kDa antibody
Signal transducer and activator of transcription 1 antibody
Signal transducer and activator of transcription 1 91kD antibody Signal transducer and activator of transcription 1-alpha/beta antibody STAT 1 antibody Stat1 antibody STAT1_HUMAN antibody STAT91 antibody Transcription factor ISGF 3 components p91 p84 antibody Transcription factor ISGF-3 components p91/p84 antibody |
|---|---|
| Molecular Weight(MW) | 87 kDa |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of Phospho-STAT1 (S727) on different lysates with Rabbit anti-Phospho-STAT1 (S727) antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane), Lane 2: Mouse brain tissue lysate (20 µg/Lane), Lane 3: Rat brain tissue lysate (20 µg/Lane), Lane 4: Mouse brain treated with λpp for 1 hour tissue lysate (20 µg/Lane). Exposure time: 3 minutes 10 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-Phospho-STAT1 (S727) antibody 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-Phospho-STAT1 (S727) antibody 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Phospho-STAT1 (S727) antibody 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 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.
ICC/IF
Immunocytochemistry analysis of HeLa cells treated with or without λpp labeling Phospho-STAT1 (S727) with Rabbit anti-Phospho-STAT1 (S727) antibody at 1/2,000 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-STAT1 (S727) antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling Phospho-STAT1 (S727). Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
WB
(B) DEF cells were transfected with a lentiviral vector expressing duck RIG-I and a lentivirus encoding ZsGreen for 24 h, and then infected with DEV at an MOI of 1 or mock-infected for 48 h, Expression levels of STAT1 and pSTAT1 in the cell lysates were analyzed by western blotting, with β-actin protein as the control.| Application Notes | WB:1:2000-1:5000 IHC1:200-1:1000ICC/IF:1:2000 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
