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생물학적 설명

특이성	β Amyloid (Aβ) 1-42 Antibody [H15J22]는 총 β Amyloid 1-42 단백질의 내인성 수준을 검출합니다.
배경	Amyloid β 1-42 (Aβ42)는 뉴런에서 β-세크레타아제와 γ-세크레타아제에 의한 순차적 절단을 통해 아밀로이드 전구체 단백질(APP)로부터 생성되는 42개의 아미노산 펩타이드입니다. 구조적으로 Aβ42는 C-말단에서 매우 소수성이어서 α-나선에서 β-병풍 구조로의 형태 변화를 촉진하여 가용성 올리고머, 원섬유 및 불용성 아밀로이드 섬유로의 응집을 용이하게 합니다. Aβ42는 주로 뇌, 특히 신피질에서 발현되며, 더 짧은 Aβ40 형태보다 응집 경향이 더 높고 신경독성이 더 강합니다. 기능적으로, Aβ42의 병리학적 축적, 특히 가용성 올리고머 형태는 시냅스 신호 전달을 방해하고 기억력을 손상시키며 플라크 형성에 기여함으로써 알츠하이머병 발병에 강력하게 연루되어 있습니다.

특이성

β Amyloid (Aβ) 1-42 Antibody [H15J22]는 총 β Amyloid 1-42 단백질의 내인성 수준을 검출합니다.

배경

Amyloid β 1-42 (Aβ42)는 뉴런에서 β-세크레타아제와 γ-세크레타아제에 의한 순차적 절단을 통해 아밀로이드 전구체 단백질(APP)로부터 생성되는 42개의 아미노산 펩타이드입니다. 구조적으로 Aβ42는 C-말단에서 매우 소수성이어서 α-나선에서 β-병풍 구조로의 형태 변화를 촉진하여 가용성 올리고머, 원섬유 및 불용성 아밀로이드 섬유로의 응집을 용이하게 합니다. Aβ42는 주로 뇌, 특히 신피질에서 발현되며, 더 짧은 Aβ40 형태보다 응집 경향이 더 높고 신경독성이 더 강합니다. 기능적으로, Aβ42의 병리학적 축적, 특히 가용성 올리고머 형태는 시냅스 신호 전달을 방해하고 기억력을 손상시키며 플라크 형성에 기여함으로써 알츠하이머병 발병에 강력하게 연루되어 있습니다.

사용 정보

응용

IHC

희석

IHC

1:1000

반응성

Mouse, Rat, Human

출처

Rabbit Monoclonal Antibody

MW

보관 완충액

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

보관
(수령일로부터)

-20°C (avoid freeze-thaw cycles), 2 years

실험 방법
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.

실험 방법

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.

참조

https://pubmed.ncbi.nlm.nih.gov/28713158/
https://pubmed.ncbi.nlm.nih.gov/23406382/

적용 데이터

IHC

Selleck 검증

Immunohistochemical analysis of formalin fixed paraffin embedded human brain tissue with F3375 at 1:1000 dilution.