Phospho-HSF1 (S326) Recombinant Rabbit Monoclonal Antibody [SU31-03]
cat.: ET1608-11
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IP
Clonality: Monoclonal
Clone number: SU31-03
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: 57 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser326 of Human HSF1 aa 301-350 / 529.
Positive control: HeLa 42℃ heat shocked for 30 minutes cell lysate, Hela, AGS, MCF-7, human breast tissue, human tonsil tissue, human lung tissue, human thyroid tissue, human skin tissue, human pancreas tissue, rat bladder tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IP
1:1,000-1:2,000
1:50-1:200
1-2μg/sample
Uniprot #: SwissProt: Q00613 Human
Alternative names: Heat shock factor 1 Heat shock factor protein 1 Heat shock transcription factor 1 HSF 1 hsf1 HSF1_HUMAN HSTF 1 HSTF1
Images
ET1608-11_1.jpg Fig1: Western blot analysis of Phospho-HSF1 (S326) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: BT-20 cell lysate
Lane 3: MCF-7 cell lysate
ET1608-11_2.jpg Fig2: Western blot analysis of Phospho-HSF1 (S326) on different lysates with Rabbit anti-Phospho-HSF1 (S326) antibody (ET1608-11) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa 42℃ heat shocked for 30 minutes cell lysate
Lane 3: HeLa 42℃ heat shocked for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 80 kDa

Exposure time: 1 minute 59 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 (ET1608-11) at 1/1,000 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.
ET1608-11_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Phospho-HSF1 (S326) antibody (ET1608-11) at 1/800 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 (ET1608-11) at 1/800 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.
ET1608-11_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-HSF1 (S326) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1608-11, 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.
ET1608-11_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-Phospho-HSF1 (S326) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1608-11, 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.
ET1608-11_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-Phospho-HSF1 (S326) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1608-11, 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.
ET1608-11_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-Phospho-HSF1 (S326) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1608-11, 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.
ET1608-11_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Phospho-HSF1 (S326) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1608-11, 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.
ET1608-11_9.jpg Fig9: Phospho-HSF1 (S326) was immunoprecipitated from 0.2 mg HeLa cell lysate with ET1608-11 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1608-11 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: ET1608-11 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of ET1608-11 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.