HSP27 [p Ser78, p Ser82, p Ser82] Antibody (1026B) [Unconjugated]

Western blot shows lysates of MCF-7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 20 mJ/cm2ultraviolet light (UV) with a 30 minute recovery. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human Phospho-HSP27 (S78/S82) Monoclonal Antibody (Catalog # MAB23141) 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.

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 Phospho-HSP27 (S78/S82) Monoclonal Antibody (Catalog # MAB23141) at a 10 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.

Phosphorylation of Akt, RSK, and CREB during T. cruzi infection. Lysates from primary HCoEpiC infected with T. cruzi at multiple time points were resolved by SDS-PAGE, blotted, and probed with antibodies against (A) p-HSP27 (B) p-Akt T308, (C) p-Akt S473, (D) p-RSK, and (E) p-CREB, and developed as described. The blots were stripped and reprobed with antibodies against the respective total proteins. The blots were developed by chemiluminescence and scanned. The normalized fold change in the level of each phosphorylated protein was determined and plotted in the bar graph below the respective blot. The bar graphs represent mean values ± SE from three independent biological replicates. The value of p<0.05 was considered significant; *p<0.05 **p<0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30222739), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clone: 1026B
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

HSP27 [p Ser78, p Ser82, p Ser82] Antibody (1026B) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Phosphopeptide containing the human HSP27 S78/S82 site
  Accession # P04792
• Modification: p Ser82, p Ser78, p Ser82
• Specificity: Detects human HSP27 when dually phosphorylated at S78/S82 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: HSPB1
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
• Buffer: 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.
• Preservative: Sodium Azide
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HSP27 [p Ser78, p Ser82, p Ser82] Antibody (1026B) [Unconjugated]

• 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

Background

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. Ser78 and Ser82 of HSP27 are phosphorylated in vivo in response to growth factors or heat shock, and the extent of phosphorylation plays a role in determining specific functions. HSP27 forms a large oligomer and 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.

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HSP27 Antibody (MAB23141)(2)

Publications using MAB23141

• Yue Zhou, Ryota Oki, Akihiro Tanaka, Leixin Song, Atsushi Takashima, Naru Hamada et al. Cellular stress induces non-canonical activation of the receptor tyrosine kinase EphA2 through the p38-MK2-RSK signaling pathway Journal of Biological Chemistry 2023-05-01 [PMID: 37059179]
• JM Santiago-O, ES Kleinerman, MG Hollomon, A Livingston, WL Wang, JW Tsai, NB Gordon Phosphorylated heat shock protein 27 as a potential biomarker to predict the role of chemotherapy-induced autophagy in osteosarcoma response to therapy Oncotarget, 2017-08-17;9(2):1602-1616. 2017-08-17 [PMID: 29416717] (Western Blot, Human)

Bioinformatics

• Gene Symbol: HSPB1
• Uniprot:
  • P04792()