PDK4 Antibody Staining Protocol for Immunohistochemistry

 

Description: PDK4 belongs to the PDK/BCKDK protein kinase subfamily. PDK4 contributes to the regulation of glucose metabolism by inhibiting the mitochondrial pyruvate dehyrogenase complex by phosphorylating the E1 alpha subunit. PDK4 is upregulated in many tissues of the body during starvation.

 

Primary Antibody

Name: PDK4 Antibody

Clone: Rabbit Polyclonal

Supplier: LifeSpan BioSciences

Catalog Number: LS-B3459

Dilution: 1:30 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 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 Chromogen Substrate
Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human, mouse
Fixation: Formalin fixed paraffin sections
Positive Control: Human adrenal, pancreas
Negative Control: Omit primary antibody, isotype control, 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

 

References: