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

R&D Systems, part of Bio-Techne | Catalog # AF2314

R&D Systems, part of Bio-Techne
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AF2314
AF2314-SP
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Flow Cytometry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

View all HSP27 Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human HSP27 S78/S82 sites

Specificity

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

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antibody

Detection of Human and Mouse Phospho-HSP27 (S78/S82) antibody by 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/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.
Phospho-HSP27 (S78/S82) antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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.
Detection of Human HSP27 by Western Blot

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.

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

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line unstimulated or stimulated with 20 mJ/cm2 ultraviolet radiation

Western Blot

0.1 µg/mL
Sample: UV-C-treated HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen and protein A Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

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.

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

  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

Alternate Names

HSP25, HSPB1

Entrez Gene IDs

3315 (Human); 15510 (Mouse); 24471 (Rat)

Gene Symbol

HSPB1

Additional HSP27 Products

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

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antibody

For research use only

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