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HSP27 Antibody Staining Protocol for Immunohistochemistry
Description: The heat-shock proteins (HSPs) belong to a larger group of polypeptides, the stress proteins, that are induced in various combinations in response to environmental challenges and developmental transitions. Synthesis of the small (27-kD) HSP has been shown to be correlated with the acquisition of thermotolerance. The deduced 199-amino acid HSP27 protein shows sequence similarity to mammalian alpha-crystallins. Approximately 20% of its residues are susceptible to phosphorylation. The HSP27 gene, which is mapped to 7q11.23 and has 3 exons1, produced a 2.2-kb transcript in an in vitro transcription assay. Decreasing ROS in cells expressing mutant huntingtin, HSP27 protects cells against oxidative stress2. In other words, HSP27 is a suppressor of polyglutamine (polyQ)-mediated cell death3. Furthermore, MAPKAPK5 is a major stress-activated kinase that can phosphorylate HSP27 in vitro.
Primary Antibody
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Name: HSP27 IHC Antibody |
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Clone: Rabbit Polyclonal |
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Supplier: IHC World |
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Catalog Number: IW-PA1121 |
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Dilution: Ready to Use |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95-100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse, rat |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Human breast cancer |
| Negative Control: Omit primary antibody, isotype control, absorption control |
| Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
References: