STAT6 Recombinant Rabbit Monoclonal Antibody [SY13-09]
cat.: ET1605-49
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IP, FC, IF-Tissue
Clonality: Monoclonal
Clone number: SY13-09
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: 94 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human STAT6 C terminal.
Positive control: Hela-si NT cell lysate, Hela-si STAT6 cell lysate, THP-1 cell lysate, Jurkat cell lysate, Raji cell lysate, HeLa cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, human solitary fibrous tumor tissue, Hela, HepG2, NIH/3T3.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IF-Tissue
  FC
  IP
1:2,000
1:200-1:500
1:3,000
1:200
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P42226 Human | P52633 Mouse
Alternative names: 12S1644 D12S1644 IL 4 STAT IL-4 Stat IL4 STAT Interleukin 4 Induced Interleukin 4 Induced Transcription Factor IL4 STAT Signal transducer and activator of transcription 6 Signal Transducer And Activator Of Transcription 6 Interleukin 4 Induced Signal Transducer And Activator Of Transcription 6 Nirs Variant 1 Signal transducer and activator of transcription 6, interleukin 4 induced STAT 6 STAT interleukin4 induced STAT, interleukin4 induced Stat6 STAT6_HUMAN STAT6B STAT6C Transcription factor IL 4 STAT
Images
ET1605-49_1.jpg Fig1: Western blot analysis of STAT6 on different lysates with Rabbit anti-STAT6 antibody (ET1605-49) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: Raji cell lysate (20 µg/Lane)
Lane 4: HeLa cell lysate (20 µg/Lane)
Lane 5: MDA-MB-231 cell lysate (20 µg/Lane)
Lane 6: NIH/3T3 cell lysate (20 µg/Lane)
Lane 7: RAW264.7 cell lysate (20 µg/Lane)

Predicted band size: 94 kDa
Observed band size: 100 kDa

Exposure time: 2 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 (ET1605-49) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1605-49_2.jpg Fig2: Western blot analysis of STAT6 on different lysates with Rabbit anti-STAT6 antibody (ET1605-49) at 1/500 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si STAT6 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 94 kDa
Observed band size: 100 kDa

Exposure time: 1 minute 34 seconds;
4-20% SDS-PAGE gel.

ET1605-49 was shown to specifically react with STAT6 in Hela-si NT cells. Weakened band was observed when Hela-si STAT6 sample was tested. Hela-si NT and Hela-si STAT6 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1605-49, 1/500) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA 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.
ET1605-49_3.jpg Fig3: STAT6 was immunoprecipitated from 0.5 mg Hela whole cell lysates with ET1605-49 at 2 μg/mL. Western blot was performed from the immunoprecipitate using ET1605-49 at 1/500 dilution for 45 minutes at room temperature. Goat anti-Rabbit IgG-HRP Secondary Antibody (HA1001) was used at 1:300,000 dilution for 30 minutes at room temperature.

Lane 1: Hela whole cell lysates at 10 μg;
Lane 2: STAT6 (ET1605-49) IP in Hela whole cell lysates;
Lane 3: Rabbit IgG instead of STAT6 (ET1605-49) in Hela whole cell lysates.

Predicted band size: 94 kDa
Observed band size: 100 kDa

Exposure time: 2 minutes;

8% SDS-PAGE gel.
ET1605-49_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human solitary fibrous tumor tissue with Rabbit anti-STAT6 antibody (ET1605-49) 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 (ET1605-49) 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.
ET1605-49_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-STAT6 antibody (ET1605-49) at 1/3,000 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 (ET1605-49) at 1/3,000 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.
ET1605-49_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-STAT6 antibody (ET1605-49) at 1/3,000 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 (ET1605-49) at 1/3,000 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.
ET1605-49_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-STAT6 antibody (ET1605-49) at 1/3,000 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 (ET1605-49) at 1/3,000 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.
ET1605-49_8.jpg Fig8: Immunocytochemistry analysis of MDA-MB-231 cells labeling STAT6 with Rabbit anti-STAT6 antibody (ET1605-49) at 1/500 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-STAT6 antibody (ET1605-49) at 1/500 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.
ET1605-49_9.jpg Fig9: Flow cytometric analysis of HeLa cells labeling STAT6.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1605-49, 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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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