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

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

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

 

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