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Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antibody

Catalog #: AF2314
3 Citations Datasheet / COA / SDS
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AF2314
AF2314-SP
Detection of Human and Mouse Phospho-HSP27 (S78/S82) by Western Blot.
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Citations (3)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antibody Summary

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 either lyophilized or 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. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human and Mouse Phospho-HSP27 (S78/S82) antibody by Western Blot. View Larger

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/m2UV-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 # 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) antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

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/cm2ultraviolet 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 # 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.

Western Blot Detection of Human HSP27 by Western Blot View Larger

Detection of Human HSP27 by Western Blot Enrichment of TICs decreases PP2A activity and increases Hsp27 activation in other solid tumors.CCS and HCW colorectal cancer cells, A549 lung cancer cells, HTB186 medulloblastoma cells and SAS, oral cancer cells were continually cultured under serum depletion in the presence of EGF (10 ng/mL) and FGF2 (10 ng/mL) to enrich TICs (E+F) or in serum-containing medium (CTR) as a control. (A) Immunoblots for pluripotent markers. (B) Immunoblots for phosphorylated PP2A (pPP2A), PP2A and Hsp27. (C) Immunoblots for caspase 9 and 3. (D) Schema demonstrates the antiapoptosis pathway of colorectal TICs in response to in vitro hypoxia and serum depletion. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0049605), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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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Citations for Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antibody

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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  1. Einfluss der Phosphorylierung des Hitzeschockproteins 27 auf das Expressionsprofil von parodontalen Ligamentfibroblasten bei mechanischer Belastung
    Authors: Agnes Schröder, Kathrin Wagner, Fabian Cieplik, Gerrit Spanier, Peter Proff, Christian Kirschneck
    Journal of Orofacial Orthopedics / Fortschritte der Kieferorthopädie
  2. A Novel Kinase Function of a Nucleoside-diphosphate-kinase Homolog in P. gingivalis is Critical in Subversion of Host Cell Apoptosis by Targeting Heat-Shock Protein 27
    Authors: J Lee, JS Roberts, KR Atanasova, N Chowdhury, Ö Yilmaz
    Cell. Microbiol., 2018-02-07;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Survival of cancer stem cells under hypoxia and serum depletion via decrease in PP2A activity and activation of p38-MAPKAPK2-Hsp27.
    Authors: Lin, Shih-Pei, Lee, Yi-Ting, Wang, Jir-You, Miller, Stephani, Chiou, Shih-Hwa, Hung, Mien-Chi, Hung, Shih-Chi
    PLoS ONE, 2012-11-20;7(11):e49605.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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