Phospho-HSF1 (Ser326) Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb559454
• Category: Antibodies
• Description: Phospho-HSF1 (Ser326) Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 37E4
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human HSF1 around the phosphorylation site of Ser326 L(p-S)PT
• Antibody Type: Recombinant Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:50-200, IHC-F=1:50-200, IF=1:50-200, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 57 kDa
• Target: HSF1
• UniProt ID: Q00613
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: HSF1(S326); HSF1(phospho S326); p-HSF1(phospho S32
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-phospho-HSF1 (Ser326) antibody (orb559454), dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Monoclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human endometrial carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Polyclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Polyclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse heart), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Polyclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse lung), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Polyclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse pancreas), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Polyclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse spleen), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-HSF1 (Ser326)) Polyclonal Antibody, Unconjugated (orb559454) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Normal human HeLa cell lysates, Lane 2: Hela cells heated at 42°C for 30 minutes, Primary: Anti-phospho-HSF1 (Ser326) (orb559454) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 57 kDa, Observed band size: 80 kDa.