Western blot analysis of various lysates using STAT1 Rabbit mAb (A19563) 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.
Chromatin immunoprecipitation was performed with 35 μg of cross-linked chromatin from HeLa cells treated by TNF-α (30ng/ml, 1h) using 5 μg of STAT1 Rabbit mAb (A19563). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of STAT1 across chromosome 6 (upper panel) and the genomic region encompassing TAP1, a representative gene enriched in STAT1 (lower panel).
Chromatin immunoprecipitation was performed with 35 μg of cross-linked chromatin from HeLa cells treated by TNF-α (30ng/ml, 1h) using 5 μg of STAT1 Rabbit mAb (A19563). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of STAT1 in the representative genomic region surrounding TAP1 gene.
Immunoprecipitation analysis of 600 μg extracts of HeLa cells using 3 μg STAT1 antibody (A19563). Western blot was performed from the immunoprecipitate using STAT1 antibody (A19563) at a dilution of 1:1000.
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.
Immunohistochemistry analysis of paraffin-embedded Rat ovary tissue using JAK1 Rabbit mAb (A11963) at a dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using JAK1 Rabbit mAb (A11963) at a dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue using JAK1 Rabbit mAb (A11963) at a dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.
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 Bufferr (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 Bufferr (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 Bufferr (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 Bufferr (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,NIH/3T3 and PC-12 cells were treated by TNF-α (20 ng/ml) at 37℃ for 30 minutes.
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: 20s.
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.
Western blot analysis of various lysates 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 Enhanced Kit (RM00021).
Exposure time: 3min.
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.
Confocal imaging of C2C12 cells using SOCS2 Rabbit mAb (A9190, dilution 1:200) followed by a further incubation with Cy3 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.
Confocal imaging of MCF7 cells using SOCS2 Rabbit mAb (A9190, dilution 1:200) followed by a further incubation with Cy3 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.
Western blot analysis of various lysates using Phospho-STAT3-S727 Rabbit mAb (AP0715) at 1:1000 dilution. NIH/3T3 and PC-12 cells were treated by TNF-α (20 ng/mL) at 37℃ for 30 minutes.
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: 120s.
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 Bufferr (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 Bufferr (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 Bufferr (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 Bufferr (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 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 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 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
Immunoprecipitation analysis of 300 μg extracts of HeLa cells using 3 μg STAT4 antibody (A4523). Western blot was performed from the immunoprecipitate using STAT4 antibody (A4523) at a dilution of 1:1000.
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.
