DNA damage repair markers

Unrepaired DNA damage leads to mutagenesis or cell death.

DNA mutations are a known factor in the tumorigenesis of many cancer types including colon and skin cancer, leukemia, lymphoma, and breast or ovarian cancer.


The accumulation of DNA damage in myocytes or neurons can result in degenerative diseases (1). The human brain consumes 20% of the body’s entire oxygen intake to generate sufficient energy to maintain its proper functioning. Since ROS (Reactive Oxygen Species) are a natural side-product of the pathways of this energy production, they can be easily accumulated in the brain and induce DNA damage.

Every day, cells are exposed to a broad range of DNA-damaging factors (2). As a safeguard for genome stability, they have developed many repair pathways, especially for damage from oxidation, hydrolysis, dimerization of DNA-bases, and single or double strand DNA breaks (Figure 1).

  

Figure 1. Overview of DNA damage-specific repair pathways. Response mechanisms: Direct reversal, MMR (MisMatch Repair), BER (Base Excision Repair), NER (Nucleotide Excision Repair), HR (Homologous Recombination), MMEJ (Microhomology-Mediated End Joining), and NHEJ (Non-Homologous End Joining).

Figure 2. Immunohistochemical analysis of paraffin-embedded human small intestine tissue slide using 60277-1-Ig ( CHK1 Antibody) at dilution of 1:50 (under 10x lens).

Climate change and increased environmental pollution are extending DNA-damaging factors. It is therefore important to understand the DNA damage repair mechanisms. Tomas Lindahl, Paul Modrich, and Aziz Sancarwere pioneers in discovering the molecular basis of DNA damage repair pathways (2015, with the Nobel Prize in Chemistry).

IHCeasy by proteintech is a ready-to-use IHC kit and it is the most complete IHC kit on the market. Primary antibody included along with other reagents.

WB result of CDC25C antibody (16485-1-AP; 1:4000; incubated at room temperature for 1.5 hours) with sh-Control and sh-CDC25C transfected HeLa cells

Figure 3. WB result of CDC25C antibody (16485-1-AP; 1:4000; incubated at room temperature for 1.5 hours) with sh-Control and sh-CDC25C transfected HeLa cells.

WB result of NBS1 antibody (55025-1-AP; 1:2000; incubated at room temperature for 1.5 hours) with sh-Control and sh-NBS1 transfected HeLa cells

Figure 4. WB result of NBS1 antibody (55025-1-AP; 1:2000; incubated at room temperature for 1.5 hours) with sh-Control and sh-NBS1 transfected HeLa cells

DNA damage repair markers

DNA damage marker

Type

Cat. No.

Damage type/

Repair mechanism

ATM

Rabbit

27156-1-AP

DSB

ATR

Rabbit

19787-1-AP

SSB

ATRIP

Rabbit

11327-1-AP

SSB

Cdc25A

Rabbit

55031-1-AP

SSB, DSB

Cdc25B

Rabbit

10644-1-AP

SSB, DSB

Cdc25C

Rabbit, Mouse

16485-1-AP  (Figure 3) ; 66912-1-Ig

SSB, DSB

Chk1

Rabbit, Mouse

25667-1-AP; 60277-1-Ig  (Figure 2)

SSB

Chk2

Rabbit

13954-1-AP

DSB

DDB1

Rabbit, Mouse

11380-1-AP; 66010-1-Ig

SSB/ NER

DDB2

Rabbit

10431-1-AP

SSB/ NER

Fen1

Rabbit

14768-1-AP

DSB/ MMEJ

H2AX

Rabbit

10856-1-AP

DSB

Ku70

Rabbit, Mouse

10723-1-AP; 66607-1-Ig

DSB/ NHEJ

Ku80

Rabbit, Mouse

16389-1-AP; 66546-1-Ig

DSB/ NHEJ

Mre11

Rabbit

10744-1-AP

DSB/ HR

Nbs1

Rabbit

55025-1-AP  (Figure 4)

DSB/ HR

PCNA

Rabbit, Mouse

10205-1-AP; 60097-1-Ig

SSB/ NER

POLD1

Rabbit

15646-1-AP

SSB

POLD2

Rabbit

10288-1-AP

SSB

POLD3

Rabbit

21935-1-AP

SSB

POLD4

Rabbit

26209-1-AP

SSB

POLE3

Rabbit

15278-1-AP

SSB

POLK

Rabbit

14455-1-AP

SSB

Rad1

Rabbit

11726-2-AP

SSB

Rad51

Rabbit, Mouse

14961-1-AP; 67024-1-Ig

DSB/ HR

RFC2

Rabbit

10410-1-AP

SSB/ NER

RFC3

Rabbit

11814-1-AP

SSB/ NER

RFC4

Rabbit

10806-1-AP

SSB/ NER

RFC5

Rabbit

10385-1-AP

SSB/ NER

RPA1

Rabbit

12448-1-AP

SSB, DSB/ HR

RPA2

Rabbit

10412-1-AP

SSB, DSB/ HR

RPA3

Rabbit

10692-1-AP

SSB, DSB/ HR

RPA4

Rabbit

18144-1-AP

SSB, DSB/ HR

 

Loading control antibodies
GAPDH Antibody mouse monoclonal GAPDH antibody WB analysis of HeLa cells
Catalog no.: 60004-1-Ig

GAPDH is commonly used as a protein loading control in western blot due to its consistently high expression in most cell types. This enzyme participates in several cellular events such as glycolysis, DNA repair, and apoptosis.

Proteintech monoclonal GAPDH antibodies are raised against a whole-protein antigen of human origin and have over 4,960 citations.

Beta Actin Antibody (KD/KO validated) WB analysis of Jurkat cells using using beta actin antibody (66009-1-Ig)
Catalog no.: 66009-1-Ig 

Beta-actin is usually used as a loading control due to its broad and consistent expression across all eukaryotic cell types and the fact that expression levels of this protein are not affected by most experimental treatments.

66009-1-Ig has been cited in over 2,460 publications and has wide species reactivity.

Homologous Recombination (HR)
Table 1. List of available antibodies against DNA damage markers and the DNA damage pathway(s)

Double Strand Breaks (DSB)

Microhomology-Mediated End Joining (MMEJ)

Nucleotide Excision Repair (NER)

Non-Homologous End Joining (NHEJ)

Single Strand Breaks (SSB)


References

1. Stein and Toiber (2017) “DNA damage and neurodegeneration: the unusual suspect”.

2. Lindahl and Barnes (2000) “Repair of endogenous DNA damage“.