HSP70 Antibody Staining Protocol for Immunohistochemistry
Description: The HSP 70 family, Heat Shock Protein, is composed of our highly conserved proteins: HSP70, HSC70, GRP75 and GRP78. These proteins serve a variety of roles: they act as molecular chaperones facilitating the assembly of multi-protein complexes, they participate in the translocation of polypeptides across cell membranes and to the nucleus and they aid in the proper folding of nascent polypeptide chains. All members of the family, except HSP70, are constitutively expressed in primate cells. HSP70 expression is strongly induced in response to heat stress. HSP70 and HSC70 play key roles in the cytosolic endoplasmic reticulum and mitochondrial import machinery and are found in both the cytosol and nucleus of mammalian cells. They are involved in the chaperoning of nascent polypeptide chains and in protecting cells against the accumulation of improperly folded proteins. Increased HSP70 levels in serum correlates with better outcome for patients after heat stroke or severe trauma. At the same time elevated serum levels of HSP70 are detected in patients with peripheral or renal vascular disease.
Primary Antibody
Name: HSP70 Antibody |
Clone: Rabbit anti-Human |
Supplier: Dako |
Catalog Number: A0500 |
Dilution: 1:300 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
Incubation Time/Temp: 60 minutes/room temperature |
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 ºC - 100 ºC/room temperature |
Heat/Cool Time: 20 minutes/20 minutes |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Cytoplasmic/nuclear |
Images: Search image |
Tissue Type: Skin |
Fixation: Formalin fixed paraffin sections |
Positive Control: Skin |
Negative Control: Omit primary antibody, isotype control or absorption control |
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
Not tested on frozen sections. |
References: