HSP27 Antibody (HSPB1/774)

Western Blot: HSP27 Antibody (HSPB1/774) [NBP2-44975] - Western Blot Analysis of HeLa cell lysate using HSP27 MAb (HSPB1/774). Image using the Azide and BSA Free format of this antibody.

Immunocytochemistry/Immunofluorescence: HSP27 Antibody (HSPB1/774) [NBP2-44975] - ICC/IF analysis of PFA-fixed MCF-7 cells labeling HSP27 with followed by Goat anti-Mouse IgG-CF488 (Green). The nuclear counterstain is Red Dot (Red)

Immunohistochemistry-Paraffin: HSP27 Antibody (HSPB1/774) [NBP2-44975] - Human Prostate Carcinoma stained with HSP27 Monoclonal Antibody (HSPB1/774)

Flow Cytometry: HSP27 Antibody (HSPB1/774) [NBP2-44975] - Flow Cytometric Analysis of PFA-fixed MCF-7 cells using HSP27 Antibody (HSPB1/774) followed by Goat anti-Mouse IgG-CF488 (Blue); Isotype Control (Red).

Immunohistochemistry-Paraffin: HSP27 Antibody (HSPB1/774) [NBP2-44975] - Human Breast Carcinoma stained with HSP27 Monoclonal Antibody (HSPB1/774)

• Reactivity: Hu, Mu, Rt, Ch, Pm, Sh
• Applications: WB, Flow, ICC/IF, IHC, IHC-P, ICC/IF
• Clone: HSPB1/774
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

HSP27 Antibody (HSPB1/774) Summary

• Immunogen: Recombinant human HSP27 protein (Uniprot: P04792)
• Localization: Predominantly cytoplasmic with some nuclear
• Specificity: It recognizes a 24-27kDa estrogen-regulated protein, identified as heat shock protein 27 (hsp27). Hsp27 was recently found to be identical to the estrogen-induced protein. About 50% of breast carcinomas are positive for hsp27 especially those that are also positive for estrogen and/or progesterone receptor. HSP27 has also been implicated in drug resistance in cancer cells.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: HSPB1
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1-2 ug/million cells
  • Immunocytochemistry/Immunofluorescence 1-2 ug/ml
  • Immunofluorescence 0.5 - 1.0 ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1-2 ug/ml
  • Western Blot 0.25-0.5 ug/ml
• Application Notes: Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
  Optimal dilution for a specific application should be determined.
• Theoretical MW: 27 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Human, Chimpanzee, Sheep, Rat, Mouse, and Chicken. Others-not known.

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein A or G purified

Notes

200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA at 1.0 mg/ml. (NBP2-48002)

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C. Antibody is stable for 24 months. Non-hazardous.

Alternate Names for HSP27 Antibody (HSPB1/774)

• 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

Hsp25 is the mouse homologue of the human Hsp27 protein, a member of the small heat shock protein family comprised of a diverse group of proteins from approx. 15 to >30kDa(1). The basic structure of most sHsps is a homologous and highly conserved amino acid sequence, with an alpha-crystallin-domain at the C-terminus and the WD/EPF domain at the less conserved N-terminus. This N-terminus is essential for the development of high molecular oligomers (2, 3). Hsp27-oligomers consist of stable dimers formed by as many as 8-40 Hsp27 protein monomers (4). The oligomerization status is connected with the chaperone activity: aggregates of large oligomers have high chaperone activity, whereas dimers have no chaperone activity (5). HSP27 is localized to the cytoplasm of unstressed cells but can redistribute to the nucleus in response to stress, where it may function to stabilize DNA and/or the nuclear membrane. It can be rapidly phosphorylated in response to physiological stimuli relevant to the cell type examined. Thus, hsp27 has been suggested to be an important intermediate in second messenger mediated signaling pathways (6). Other functions include chaperone activity (as mentioned above), thermotolerance in vivo, inhibition of apoptosis, and signal transduction. Specifically, in vitro, it acts as an ATP-independent chaperone by inhibiting protein aggregation and by stabilizing partially denatured proteins, which ensures refolding of the HSP70 complex. Hsp27 is also involved in the apoptotic signaling pathway because it interferes with the activation of cytochrome c/Apaf-1/dATP complex, thereby inhibiting the activation of procaspase-9. It is also hypothesized that hsp27 may serve some role in cross-bridge formation between actin and myosin (7). And finally, Hsp27 is also thought to be involved in the process of cell differentiation. The up-regulation of Hsp27 correlates with the rate of phosphorylation and with an increase of large oligomers. It is possible that Hsp27 may play a crucial role in termination of growth (8).

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.

Bioinformatics

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