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

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 ºC - 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic/nuclear
Images: Search image

Additional Information:

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.