專一性佳之8-oxo-dG單株抗體。Trevigen 4354-MC-050。可供為ELISA、 immunohistochemistry及immunocytochemistry之應用。－太鼎生物科技/太鼎食安科技

專一性佳之8-oxo-dG 單株抗體 (4345-MC-50)-Trevigen

造成 DNA 損傷：最為人所熟知的DNA損傷指的是DNA的鹼基被ROS氧化， DOX會經由氧化還原過程產生的自由基，例如 OH ‧ 會氧化DNA鹼基產生8-oxo-dG 和 8-oxo-dA。

8 - 羥基-2'-脫氧鳥苷（8-羰基-dG）為修飾核苷酸。為最常見的研究和檢測指標；例如, 氧化自由基造成的DNA損傷、炎症、癌變、帕金森的和阿爾茨海默氏症的相關疾病。8 - oxo-dG可以作為生理和環境中破壞DNA的一個敏感指標。Trevigen提供專一性佳之8-oxo-dG單株抗體, (4354-MC-050), 可供為ELISA, immunohistochemistry, or by immunocytochemistry之應用。

Catalog #	Product Name	Size
4354-MC-050	Anti-8-oxo-dG Monoclonal Antibody (clone 2E2)	50 µl

APPLICATIONS :

ELISA
Immunocytochemistry
Immunohistochemistry

8-hydroxy-2'-deoxyguanosine (8-oxo-dG) is a modified nucleoside, which is the most commonly studied and detected by-product of DNA damage, caused by oxidative radicals associated with inflammation, carcinogenesis, Parkinson's and Alzheimer's diseases, and also aging. 8-oxo-dG can serve as a sensitive indicator of physiological and environmental damage to DNA. This mouse monoclonal antibody is provided to enable the detection of 8-oxo-dG by ELISA, immunohistochemistry, or by immunocytochemistry.

IMMUNOGEN:

8-oxo-dG-conjugated-KLH

SOURCE:

Mouse

SPECIFICITY:

This mouse monoclonal antibody specifically binds to 8-hydroxy-2'- deoxyguanosine within DNA

ISOTYPE:

IgG2b.

PREPARATION:

This antibody is provided as purified immunoglobulin from mouse ascites in 1X PBS containing

0.01% sodium azide.

STORAGE:

Freeze in working aliquots at -20°C in a manual defrost freezer to avoid repeated freeze-thawings.

REFERENCES:

1. Soultanakis RP, Melamede RJ, Bespalov IA, Wallace SS, Beckman KB, Ames BN, Taatjes DJ, Janssen-Heininger YMW. (2000) Flourescence detection of 8-oxoguanine in nuclear and mitochondrial DNA of cultured cells using a recombinant Fab and confocal scanning laser microscopy. Free Rad Biol Med 28:987-998.

Catalog #	Product Name	Size
4354-MC-050	Anti-8-oxo-dG Monoclonal Antibody (clone 2E2)	50 µl

Anti-8-oxo-dG Monoclonal Antibody (clone 2E2)
Catalog #: 4354-MC-050

Description: This mouse monoclonal antibody specifically binds to 8-hydroxy -2’ -deoxyguanosine within DNA in H2O2-treated cells. It can be used to detect oxidative damage by ELISA (cat# 4370-096-K) and immunocytochemistry. Sufficient antibody is provided for approximately 50 slides, when a 1:250 dilution is used.

Physical state: This antibody is provided as purified immunoglobulin from mouse ascites at 0.5 mg/ml in 1X TBS containing 0.1% sodium azide.

Ig Class: IgG 2a

Storage Conditions: This antibody can be stored at -20°C or -80°C . Avoid repeated

freeze-thawing by aliquoting into smaller portions.

Applications: Immunodetection of 8-oxo-dG by ELISA, immunocytochemistry, and immunofluorescence.

Empirical determination will be required for optimal results. For optimal

outcomes, cells should be grown on a surface that allows for fixation and direct labeling

such as sterile chamber slides and coverslips. Alternatively, paraffin-embedded samples

may be used.

Immunocytochemistry Protocol:

1. Plate cells 5x 104 cells (sub-confluent) on cover slips or chamber slides o/n

2. Aspirate medium, wash cells with 1X PBS, and treat with 300μl of 100-300μM H2O 2 in 1X PBS, on ice for 20 minutes. (Be sure to establish untreated controls.)

3. Wash 3x with 1X PBS, and fix with -20°C MeOH followed by -20°C acetone at -20°C for

15 minutes each. Alternatively, cells may be fixed with 1:1 MeOH, acetone for 20minutes at -20°C . Allow to air dry.

4. Treat fixed cells with 0.05N HCl for 5 minutes on ice.

5. Wash 3x with 1X PBS, 5 minutes each.

6. Incubate with 250μl of 100μg/ml RNAse in 150mM NaCl, 15mM sodium citrate for 1hour at 37°C .

7. Wash sequentially in 1X PBS, 35%, 50% and 75% EtOH, for 3 minutes each.

8. Denature DNA in situ with 250μl 0.15N NaOH in 70% EtOH for 4 minutes.

9. Wash briefly 2x with 1X PBS.

10. Use 0.2 μg/ml (250μl) Hoechst 33342 (Immunochemistry Technologies, LLC) in 1XPBS to stain DNA for 10 minutes.

11. Wash sequentially in 70% EtOH containing 4% v/v formaldehyde, 50% and 35% EtOH, and 1X PBS for 2 minutes each.

12. Incubate in 250μl of 5μg/ml proteinase K in 20mM Tris, 1mM EDTA, pH 7.5 (TE) for10 minutes at 37°C .

13. Wash several times with 1X PBS.

14. Block non-specific binding with 5% normal goat serum in 1X PBS, 1hour at RT.

15. Wash 3x with 1X PBS, and incubate with 250μl anti-8-hydroxyguanine antibody at a concentration of 1:250 diluted in 1X PBS containing 1% BSA, 0.01% Tween 20 at 4°C o/n in a humidified chamber.