HSP27 Antibody (967060) [Unconjugated]

Western blot shows lysates of MCF-7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Human HSP27 Monoclonal Antibody (Catalog # MAB15801) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). 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 1.

HSP27 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rat Anti-Human HSP27 Monoclonal Antibody (Catalog # MAB15801) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HSP27 was detected in immersion fixed paraffin-embedded sections of human ovarian cancer tissue using Rat Anti-Human HSP27 Monoclonal Antibody (Catalog # MAB15801) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF
• Clone: 967060
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

HSP27 Antibody (967060) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human HSP27
  Met1-Lys205
  Accession # P04792
• Specificity: Detects human HSP27 in direct ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• 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.
• 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 Antibody (967060) [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 (MAB15801)(1)

Publication using MAB15801

• Guangxin Li Comparative analysis of the interaction of HSPs in dendritic cells, macrophages, RGM-1 cells infected by Helicobacter pylori Am J Transl Res, 2016-10-15;8(10):4184-4194. 2016-10-15 [PMID: 27830002] (Western Blot, Rat)

Bioinformatics

• Uniprot:
  • P04792()