Human/Mouse HSP27 Antibody

(3 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects recombinant and endogenous human and mouse HSP27 in Western blots.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human HSP27
    Accession # P04792
  • 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
  • Simple Western
    0.5 µg/mL
    See below
  • Knockout Validated
    HSP27 is specifically detected in HeLa humancervical epithelial carcinoma parental cell line but is not detectable inHSP27 knockout HeLa cell line.
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/Mouse HSP27 by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line, HeLa human cervical epithelial carcinoma cell line, and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HSP27 at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Human HSP27 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for HSP27 at approximately 31 kDa (as indicated) using 0.5 µg/mL of Rabbit Anti-Human/Mouse HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Detection of Mouse HSP27 by Simple WesternTM. Simple Western lane view shows lysates of C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for HSP27 at approximately 31 kDa (as indicated) using 0.5 µg/mL of Rabbit Anti-Human/Mouse HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Knockout Validated
Western Blot Shows Human HSP27Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLacell line, but is not detectable in knockout HeLa cell line. GAPDH(Catalog # MAB5718) is shown as a loading control.. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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, also known as HSPB1, 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. Full length human HSP27 shares 83% and 81% aa identity with mouse and rat HSP27, respectively.
  • 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Heat shock protein 27 differentiates tolerogenic macrophages that may support human breast cancer progression.
    Authors: Banerjee S, Lin CF, Skinner KA, Schiffhauer LM, Peacock J, Hicks DG, Redmond EM, Morrow D, Huston A, Shayne M, Langstein HN, Miller-Graziano CL, Strickland J, O'Donoghue L, De AK
    Cancer Res., 2011;71(2):318-27.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  2. The heat shock response and chaperones/heat shock proteins in brain tumors: surface expression, release, and possible immune consequences.
    Authors: Graner MW, Cumming RI, Bigner DD
    J. Neurosci., 2007;27(42):11214-27.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Phosphatidylinositol 3-kinase/Akt plays a role in sphingosine 1-phosphate-stimulated HSP27 induction in osteoblasts.
    Authors: Takai S, Tokuda H, Matsushima-Nishiwaki R, Hanai Y, Kato K, Kozawa O
    J. Cell. Biochem., 2006;98(5):1249-56.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
Isotype Controls
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Normal Rabbit IgG Control

Ctrl AB-105-C 43  
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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL004 10
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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Rabbit IgG APC-conjugated Antibody

Flow F0111 1  
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Rabbit IgG PE-conjugated Antibody

Flow F0110 2  
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Rabbit IgG Fluorescein-conjugated Antibody

Flow F0112 5  
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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG Antibody

WB AF008 1
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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