ET1603-40

Phospho-STAT3 (Y705) Recombinant Rabbit Monoclonal Antibody [SZ43-01]

cat.: ET1603-40

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IP
Clonality:	Monoclonal
Clone number:	SZ43-01

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: 88 kDa
Isotype:	IgG
Immunogen:	Synthetic phospho-peptide corresponding to residues surrounding Tyr705 of human STAT3.
Positive control:	HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate, Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate, A431 treated with 100ng/mL EGF for 30 minutes whole cell lysate, human lung tissue, Hela cell lysate, SK-Br-3 cell lysate, human kidney tissue, mouse skeletal muscle tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IHC-P IP	1:5,000 1:50-1:200 Use at an assay dependent concentration.
Uniprot #:	SwissProt: P40763 Human \| P42227 Mouse \| P52631 Rat
Alternative names:	1110034C02Rik Acute Phase Response Factor Acute-phase response factor ADMIO APRF AW109958 DNA binding protein APRF FLJ20882 HIES MGC16063 Signal transducer and activator of transcription 3 (acute phase response factor) Signal transducer and activator of transcription 3 STAT 3 Stat3 STAT3_HUMAN

Images

Fig1: Western blot analysis of Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa whole cell lysate
Lane 2: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate
Lane 3: Jurkat whole cell lysate
Lane 4: Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes whole cell lysate
Lane 5: A431 whole cell lysate
Lane 6: A431 treated with 100ng/mL EGF for 30 minutes whole cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 88 kDa
Observed band size: 88 kDa

Exposure time: 35 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 (ET1603-40) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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: Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) 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 (ET1603-40) 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.

Fig3: Western blot analysis of Phospho-STAT3(Tyr705) on Hela cell lysates.

Lane 1 : Hela cells, whole cell lysate, 10ug/lane
Lane 2 : Hela cells starved for 16 hours, whole cell lysates, 10ug/lane
Lane 3/4 : Hela cells starved for 16 hours, then treated with 50 ng/ml IFN alpha for 30 minutes, whole cell lysates, 10ug/lane
Lane 5 : Hela cells starved for 16 hours and treated with 50 ng/ml IFN alpha for 30 minutes, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane

All lanes :
Anti-Phospho-STAT3(Tyr705) antibody (ET1603-40) at 1:500 dilution. Anti-STAT3 antibody (ET1607-38) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution.

Predicted band size: 88 kDa
Observed band size: 88 kDa

Blocking and diluting buffer: 5% BSA.

Exposure time: 1 minute 2 seconds

Fig4: Western blot analysis of Phospho-STAT3 (Y705) on SK-Br-3 cell lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 88 kDa
Observed band size: 88 kDa

Exposure time: 2 minutes;

8% 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 (ET1603-40) at 1/500 dilution was used in 5% NFDM/TBST 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.

Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) 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 (ET1603-40) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue using anti-Phospho-STAT3 (Y705) antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-40, 1/50) for 30 minutes 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.

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