Human Phospho-HSP27 (S78/S82) Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human HSP27 when dually phosphorylated at S78/S82 in Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1026B
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen

    Phosphopeptide containing the human HSP27 S78/S82 site

    Accession # P04792

  • Formulation
    Supplied as a 0.2 μm filtered solution in PBS containing BSA, Glycerol and Sodium Azide. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1:1000 dilution
    See below
  • Immunocytochemistry
    1:100 dilution
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples

Detection of Human Phospho-HSP27 (S78/S82) by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 20 mJ/cm2 ultraviolet light (UV) with a 30 minute recovery. PVDF membrane was probed with 1:1000 dilution of Rabbit Anti-Human/Mouse/Rat Phospho-HSP27 (S78/S82) Monoclonal Antibody (Catalog # MAB23141) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-HSP27 (S78/S82) at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry
Phospho-HSP27 (S78/S82) in HeLa Human Cell Line. HSP27 phosphorylated at S78/S82 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line unstimulated (lower panel) or stimulated with 20 mJ/cm2 ultraviolet radiation (upper panel) using Rabbit Anti-Human/Mouse/Rat Phospho-HSP27 (S78/S82) Monoclonal Antibody (Catalog # MAB23141) at a 1:100 dilution for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    • 12 months from date of receipt, -20 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after opening.
    • 6 months, -20 °C under sterile conditions after opening.
Background: HSP27

Heat shock proteins (HSPs) are a family of highly conserved stress response proteins. Heat shock proteins function primarily as molecular chaperones by facilitating the folding of other cellular proteins, preventing protein aggregation or targeting improperly folded proteins to specific degradative pathways. HSPs are typically expressed at low levels under normal physiological conditions but are dramatically up-regulated in response to cellular stress. Elevated levels of HSPs have been observed in association with ischemia/reperfusion, cancer, and chronic heart failure. HSP27 is a member of the small heat shock protein family, which also includes HSP25 and the alpha -crystallins. Ser78 and Ser82 of HSP27 are phosphorylated in vivo in response to growth factors or heat shock, and the extent of phosphorylation plays a role in determining specific functions. HSP27 forms a large oligomer and functions as an anti-apoptotic molecule, regulating apoptosis through direct interaction with key components of the apoptotic pathway. HSP27 binds and sequesters cytochrome c released from the mitochondria in response to an apoptotic stimulus. This prevents the proper assembly of the apoptosome and subsequently, the activation of procaspase-9 and procaspase-3.

  • References:
    1. Gusev, N.B. et al. (2002) Biochemistry (Moscow) 67:511.
    2. Garrido, C. et al. (2001) Biochem. Biophys. Res. Commun. 286:433.
    3. Garrido, C. (2002) Cell Death Diffr. 9:483.
    4. Brvey, J-M. et al. (2000) Nat. Cell Biol. 2:645.
  • Long Name:
    Heat Shock Protein 27
  • Entrez Gene IDs:
    3315 (Human); 15510 (Mouse); 24471 (Rat)
  • Alternate Names:
    28 kDa heat shock protein; DKFZp586P1322; Estrogen-regulated 24 kDa protein; Heat shock 27 kDa protein; heat shock 27kD protein 1; heat shock 27kDa protein 1; heat shock protein beta-1; HMN2B; HS.76067; HSP25; HSP27; HSP27HSP 27; HSP28CMT2F; HSPB1; SRP27; Stress-responsive protein 27
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