8-Hydroxy-2'-deoxyguanosine Antibody [K11E8]

Aplicación: Reactividad:Species independent

Immunohistochemical analysis of formalin fixed paraffin embedded human colorectal cancer tissue with F1714 at 1:50 dilution.

Información de uso

Dilución
IHC1:100 - 1:200

Aplicación
IHC-P

Reactividad
Species independent

Fuente
Mouse Monoclonal Antibody

Tampón de almacenamiento
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Almacenamiento (desde la fecha de recepción)
-20°C (avoid freeze-thaw cycles), 2 years

Control positivo	Human colon, Epidermis from hairless mouse
Control negativo

Métodos experimentales

IHC
Experimental Protocol： Deparaffinization/Rehydration 1. Deparaffinize/hydrate sections: 2. Incubate sections in three washes of xylene for 5 min each. 3. Incubate sections in two washes of 100% ethanol for 10 min each. 4. Incubate sections in two washes of 95% ethanol for 10 min each. 5. Wash sections two times in dH2O for 5 min each. 6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min. Staining 1. Wash sections in dH2O three times for 5 min each. 2. Incubate sections in 3% hydrogen peroxide for 10 min. 3. Wash sections in dH2O two times for 5 min each. 4. Wash sections in wash buffer for 5 min. 5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature. 6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C. 7. Remove antibody solution and wash sections with wash buffer three times for 5 min each. 8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature. 9. Wash sections three times with wash buffer for 5 min each. 10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use. 11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity. 12. Immerse slides in dH2O. 13. If desired, counterstain sections with hematoxylin. 14. Wash sections in dH2O two times for 5 min each. 15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each. 16. Mount sections with coverslips and mounting medium.

IHC

Experimental Protocol：

Deparaffinization/Rehydration

1. Deparaffinize/hydrate sections:

2. Incubate sections in three washes of xylene for 5 min each.

3. Incubate sections in two washes of 100% ethanol for 10 min each.

4. Incubate sections in two washes of 95% ethanol for 10 min each.

5. Wash sections two times in dH2O for 5 min each.

6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

Staining

1. Wash sections in dH2O three times for 5 min each.

2. Incubate sections in 3% hydrogen peroxide for 10 min.

3. Wash sections in dH2O two times for 5 min each.

4. Wash sections in wash buffer for 5 min.

5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.

6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.

7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.

8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.

9. Wash sections three times with wash buffer for 5 min each.

10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.

11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.

12. Immerse slides in dH2O.

13. If desired, counterstain sections with hematoxylin.

14. Wash sections in dH2O two times for 5 min each.

15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.

16. Mount sections with coverslips and mounting medium.

Datasheet & SDS

Descripción biológica

Especificidad
8-Hydroxy-2'-deoxyguanosine Antibody [K11E8] detects both endogenous and exogenous 8-hydroxy-2'-deoxyguanosine (8-OH-dG).

Clon
K11E8

Sinónimo
8-OH-dG

Antecedentes
8-Hydroxy-2'-deoxyguanosine (8-OHdG) is a modified purine 2'-deoxyribonucleoside formed by the hydroxylation of the guanine base at the C8 position in DNA, typically resulting from oxidative damage caused by reactive oxygen species (ROS). Structurally, it consists of a deoxyguanosine molecule with a hydroxyl group at the 8th carbon of the guanine ring. 8-OHdG is widely recognized as a key biomarker of oxidative stress due to its high prevalence among oxidative DNA lesions. It accumulates in both nuclear and mitochondrial DNA and can be measured in various biological samples, including urine, tissues, and leukocyte DNA. Functionally, 8-OHdG serves as an indicator of endogenous DNA damage and has been extensively used to assess the risk and progression of cancer, aging, and degenerative diseases. Its presence reflects cellular exposure to oxidative agents and is instrumental in evaluating the impact of environmental and lifestyle factors on genomic stability.

Antecedentes

8-Hydroxy-2'-deoxyguanosine (8-OHdG) is a modified purine 2'-deoxyribonucleoside formed by the hydroxylation of the guanine base at the C8 position in DNA, typically resulting from oxidative damage caused by reactive oxygen species (ROS). Structurally, it consists of a deoxyguanosine molecule with a hydroxyl group at the 8th carbon of the guanine ring. 8-OHdG is widely recognized as a key biomarker of oxidative stress due to its high prevalence among oxidative DNA lesions. It accumulates in both nuclear and mitochondrial DNA and can be measured in various biological samples, including urine, tissues, and leukocyte DNA. Functionally, 8-OHdG serves as an indicator of endogenous DNA damage and has been extensively used to assess the risk and progression of cancer, aging, and degenerative diseases. Its presence reflects cellular exposure to oxidative agents and is instrumental in evaluating the impact of environmental and lifestyle factors on genomic stability.

Referencias
https://pubmed.ncbi.nlm.nih.gov/19412858/

Tabla de referencia para seleccionar las especificaciones del tamaño de poro de la membrana PVDF

Tamaño de la proteína	Tamaño de poro de la membrana de transferencia PVDF
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Soporte técnico

Instrucciones de manipulación

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Tamaño de la proteína	Porcentaje de gel de separación
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%