ET1607-61

STAT6 Recombinant Rabbit Monoclonal Antibody [SY02-72]

cat.: ET1607-61

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	SY02-72

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 aa 800 to the C-terminus.
Positive control:	Raji cell lysate, RAW264.7 cell lysate, mouse kidney tissue, rat kidney tissue, human kidney tissue, mouse lung tissue, mouse stomach tissue, RAW264.7, Hela, AGS, NIH/3T3.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IP FC	1:2,000 1:50 1:50 1:50-1:200 Use at an assay dependent concentration. 1:1,000
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

Fig1: Western blot analysis of STAT6 on different lysates with Rabbit anti-STAT6 antibody (ET1607-61) at 1/2,000 dilution.

Lane 1: Raji cell lysate
Lane 2: RAW264.7 cell lysate

Lysates/proteins at 30 µg/Lane.

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

Exposure time: 3 minutes; ECL: K1801;

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 (ET1607-61) at 1/2,000 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.

Fig2: Western blot analysis of STAT6 on different lysates with Rabbit anti-STAT6 antibody (ET1607-61) 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: 94 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

ET1607-61 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 (ET1607-61, 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 1:200,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-STAT6 antibody (ET1607-61) at 1/200 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 (ET1607-61) at 1/200 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-STAT6 antibody (ET1607-61) at 1/200 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 (ET1607-61) at 1/200 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 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 (ET1607-61) at 1/50 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-STAT6 antibody (ET1607-61) at 1/20 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 (ET1607-61) at 1/20 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 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 (ET1607-61) at 1/50 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.
	Fig8: Flow cytometric analysis of RAW264.7 cells labeling STAT6. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1607-61, 1/1,000) (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).

	Fig9: Immunocytochemistry analysis of Hela cells labeling STAT6 with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 dilution in 2% negative goat serum 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.
	Fig10: Immunocytochemistry analysis of AGS cells labeling STAT6 with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 dilution in 2% negative goat serum 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.
	Fig11: Immunocytochemistry analysis of NIH/3T3 cells labeling STAT6 with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-STAT6 antibody (ET1607-61) at 1/50 dilution in 2% negative goat serum 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.