Phospho-HSP27 (S78/S82) Affinity Purified Polyclonal Ab 50 UG

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Phospho-HSP27 (S78/S82) Affinity Purified Polyclonal Ab 50 UG信息二维码

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产品介绍

    基本参数

    详细说明

    • Species Reactivity

      Human, Mouse, Rat

    • Specificity

      Detects human HSP27 when dually phosphorylated at S78/S82, and mouse and rat HSP27 phosphorylated at S86.

    • Source

      Polyclonal Rabbit IgG

    • Purification

      Antigen and protein A Affinity-purified

    • Immunogen

      Phosphopeptide containing human HSP27 S78/S82 sites

    • Formulation

      Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

    • Label

      Unconjugated

    Applications

    • Recommended    
      Concentration

      Sample

    • Western Blot

      0.1 µg/mL

      See below


    • Immunocytochemistry

      5-15 µg/mL

      See below


    Please Note: Optimal dilutions should be determined by each laboratory for each application.  are available in the Technical Information section on our website.

    Data Examples

    Western Blot      
         

    Detection of Human and Mouse Phospho-HSP27 (S78/S82) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast cell line untreated (-) or treated (+) with 100 J/m2 UV-C for 30 minutes. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2314), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # ). 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 .

    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) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2314) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # ) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for .

    Preparation and Storage

    • Reconstitution

      Reconstitute at 0.2 mg/mL in sterile PBS.

    • Shipping

      The product is shipped at ambient temperature. 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

      Use a manual defrost freezer and avoid repeated freeze-thaw cycles.    

      • 12 months from date of receipt, -20 to -70 °C as supplied.

      • 1 month, 2 to 8 °C under sterile conditions after reconstitution.

      • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

    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. HSP27 forms a large oligomer and the extent of phosphorylation plays a role in determining specific functions. HSP27 also 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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