Thymine Dimer Antibody Staining Protocol for Immunohistochemistry


Description: Thymine dimers are the most dominant form of chemical modification that occur on DNA after exposure to UV-light. It detects thymine dimers in single-stranded DNA, and has a lower affinity for the dimer in short oligonucleotides and is well suited for anyone working in the field of DNA damage and repair.


Primary Antibody

Name: Thymine Dimer Antibody

Clone: Mouse monoclonal

Supplier: Affitech

Catalog Number: 1000-1

Dilution: 1:2000 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
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Nuclear
Images: Search image

Additional Information:
Species Reactivity: Human, mouse, Swine
Fixation: Formalin fixed paraffin sections. Not suitable for frozen sections.
Positive Control: Skin (UV exposed)
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