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

Catalog #: AF2314
4 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF2314
AF2314-SP

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

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Citations (4)

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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

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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

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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

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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

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Gusev, N.B. et al. (2002) Biochemistry (Moscow) 67:511.
Garrido, C. et al. (2001) Biochem. Biophys. Res. Commun. 286:433.
Garrido, C. (2002) Cell Death Diffr. 9:483.
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

Product Datasheets

Product Datasheet

COA

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Related Research Areas

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.

4 Citations: Showing 1 - 4
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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
Multiscale networks in multiple sclerosis
Authors: Kennedy, KE;Kerlero de Rosbo, N;Uccelli, A;Cellerino, M;Ivaldi, F;Contini, P;De Palma, R;Harbo, HF;Berge, T;Bos, SD;Høgestøl, EA;Brune-Ingebretsen, S;de Rodez Benavent, SA;Paul, F;Brandt, AU;Bäcker-Koduah, P;Behrens, J;Kuchling, J;Asseyer, S;Scheel, M;Chien, C;Zimmermann, H;Motamedi, S;Kauer-Bonin, J;Saez-Rodriguez, J;Rinas, M;Alexopoulos, LG;Andorra, M;Llufriu, S;Saiz, A;Blanco, Y;Martinez-Heras, E;Solana, E;Pulido-Valdeolivas, I;Martinez-Lapiscina, EH;Garcia-Ojalvo, J;Villoslada, P;
PLoS computational biology
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry
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
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