Western blot analysis of various lysates using STAT1 Rabbit mAb (A19563) at 1:20000 dilution incubated at room temperature for 1.5 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 20s.
Western blot analysis of various lysates using STAT1 Rabbit mAb (A19563) at 1:20000 dilution incubated at room temperature for 1.5 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.
Confocal imaging of HeLa cells (treated with hIFN-α1) and HeLa cells (untreated) using STAT1 Rabbit mAb (A19563, dilution 1:600) followed by a further incubation with Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
Immunoprecipitation of STAT1 from 400 µg extracts of HeLa cells was performed using 0.5 µg of STAT1 Rabbit mAb (A19563). Rabbit Control IgG (AC005) was used to precipitate the Control IgG sample. IP samples were eluted with 1x Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using STAT1 Rabbit mAb (A19563) at a dilution of 1:20000.
Chromatin immunoprecipitation was performed with 15 μg of cross-linked chromatin from RAW 264.7 cells were treated with IFN-γ (50ng/ml, 30min), using 0.5 μg of STAT1 Rabbit mAb(A19563) and Rabbit IgG isotype control (AC042). The enrichment of immunoprecipitated DNA at different genomic loci was examined by quantitative PCR. The histogram compares the ratio of the immunoprecipitated DNA to the input at given loci.
Confocal imaging of frozen sections of Mouse thymus using STAT1 Rabbit mAb (A19563, dilution 1:600) followed by a further incubation with Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Microwave antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.
Confocal imaging of frozen sections of Rat spleen tissue using STAT1 Rabbit mAb (A19563, dilution 1:200) followed by a further incubation with Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Microwave antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.
Western blot analysis of lysates from Mouse lung using JAK1 Rabbit mAb (A11963) at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Enhanced Kit (RM00021).
Exposure time: 180s.
Western blot analysis of lysates from Raji cells, using SHP1 Rabbit mAb (A19111) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.
Western blot analysis of various lysates using SHP1 Rabbit mAb (A19111) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 3min.
Immunohistochemistry analysis of paraffin-embedded Human spleen tissue using SHP1 Rabbit mAb (A19111) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue using SHP1 Rabbit mAb (A19111) at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using SHP1 Rabbit mAb (A19111) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using SHP1 Rabbit mAb (A19111) at a dilution of 1:800 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunofluorescence analysis of paraffin-embedded rat spleen using SHP1 Rabbit mAb (A19111) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
Immunofluorescence analysis of paraffin-embedded mouse spleen using SHP1 Rabbit mAb (A19111) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
Western blot analysis of various lysates using Phospho-STAT1-Y701 Rabbit mAb (AP0054) at 1:1000 dilution incubated overnight at 4℃. HeLa cells, NIH/3T3 cells and PC-12 cells were treated with TNF-α (100 ng/mL) at 37℃ for 30 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 90s.
Western blot analysis of lysates from K-562 cells, using SOCS2 Rabbit mAb (A9190) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1s.
Immunohistochemistry analysis of paraffin-embedded Rat lung using SOCS2 Rabbit mAb (A9190) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.
Western blot analysis of various lysates using Phospho-STAT3-S727 Rabbit mAb (AP0715) at 1:10000 dilution incubated overnight at 4℃. HeLa cells and NIH/3T3 cells were treated with λPP (2 U/μL) at 37℃ for 1 hour.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 45 s.
Immunohistochemistry analysis of paraffin-embedded Mouse kidney using Phospho-STAT3-S727 Rabbit mAb (AP0715) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Mouse liver using Phospho-STAT3-S727 Rabbit mAb (AP0715) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Rat colon using Phospho-STAT3-S727 Rabbit mAb (AP0715) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Rat liver using Phospho-STAT3-S727 Rabbit mAb (AP0715) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate buffer (pH 6.0) prior to IHC staining.
Western blot analysis of various lysates using STAT4 Rabbit mAb (A4523) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 90s.
Immunofluorescence analysis of C6 cells using STAT4 Rabbit mAb (A4523) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
Immunofluorescence analysis of NIH-3T3 cells using STAT4 Rabbit mAb (A4523) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
Western blot analysis of various lysates using STAT6 Rabbit mAb (A19120) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 3min.
Immunofluorescence analysis of HeLa cells using STAT6 Rabbit mAb (A19120) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
