Phospho-HSP27 (Ser15) Rabbit Polyclonal Antibody

• Catalog Number: orb5483
• Category: Antibodies
• Description: Phospho-HSP27 (Ser15) Rabbit Polyclonal Antibody
• Target: HSPB1
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Predicted Reactivity: Bovine, Sheep
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human HSP27 around the phosphorylation site of Ser15 GP(p-S)WD
• UniProt ID: P04792
• RRID: AB_10934380
• MW: 23 kDa
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg /Test
• Antibody Type: Primary Antibody
• 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.
• Alternative names: Hsp27(phospho S15); p-Hsp27(phospho S15); HSP27(ph
• Note: For research use only

Blank control (blue line): A431 cells (fixed with 70% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C). Primary Antibody (green line): Rabbit Anti-Phospho-HSP27 (Ser15) antibody (orb5483), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 1 µg/Test.

Hela cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Phospho-HSP27 (Ser15)) polyclonal Antibody, Unconjugated (orb5483) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (human gastric 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 (P-HSP27 (Ser15)) Polyclonal Antibody, Unconjugated (orb5483) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Lane 1: MCF-7 (Human) Cell Lysate at 30 ug, Lane 2: U251 (Human) Cell Lysate at 30 ug, Primary: Anti-Phospho-HSP27 (Ser15) (orb5483) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 27 kD, Observed band size: 27 kD.