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Biologische Beschrijving

Specificiteit	β Amyloid (Aβ) 1-42 Antibody [H15J22] detecteert endogene niveaus van totaal β Amyloid 1-42 eiwit.
Achtergrond	Amyloid β 1-42 (Aβ42) is een peptide van 42 aminozuren dat wordt gegenereerd uit het amyloïde prekursorproteïne (APP) via sequentiële splitsing door β-secretase en γ-secretase in neuronen. Structureel is Aβ42 sterk hydrofoob aan de C-terminus, wat een conformationele verschuiving van α-helix naar β-sheet bevordert die aggregatie in oplosbare oligomeren, protofibrillen en onoplosbare amyloïde fibrillen vergemakkelijkt. Aβ42 wordt voornamelijk tot expressie gebracht in de hersenen, vooral in de neocortex, en is meer aggregatiegevoelig en neurotoxisch dan de kortere Aβ40-vorm. Functioneel is pathologische accumulatie van Aβ42 – met name als oplosbare oligomeren – sterk betrokken bij de pathogenese van de ziekte van Alzheimer door synaptische signalering te verstoren, het geheugen aan te tasten en bij te dragen aan plaquevorming.

Specificiteit

β Amyloid (Aβ) 1-42 Antibody [H15J22] detecteert endogene niveaus van totaal β Amyloid 1-42 eiwit.

Achtergrond

Amyloid β 1-42 (Aβ42) is een peptide van 42 aminozuren dat wordt gegenereerd uit het amyloïde prekursorproteïne (APP) via sequentiële splitsing door β-secretase en γ-secretase in neuronen. Structureel is Aβ42 sterk hydrofoob aan de C-terminus, wat een conformationele verschuiving van α-helix naar β-sheet bevordert die aggregatie in oplosbare oligomeren, protofibrillen en onoplosbare amyloïde fibrillen vergemakkelijkt. Aβ42 wordt voornamelijk tot expressie gebracht in de hersenen, vooral in de neocortex, en is meer aggregatiegevoelig en neurotoxisch dan de kortere Aβ40-vorm. Functioneel is pathologische accumulatie van Aβ42 – met name als oplosbare oligomeren – sterk betrokken bij de pathogenese van de ziekte van Alzheimer door synaptische signalering te verstoren, het geheugen aan te tasten en bij te dragen aan plaquevorming.

Gebruiksinformatie

Toepassing

IHC

Verdunning

IHC

1:1000

Reactiviteit

Mouse, Rat, Human

Bron

Rabbit Monoclonal Antibody

MW

Opslagbuffer

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

Opslag
(Vanaf de datum van ontvangst)

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

Experimentele Methoden
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.

Experimentele Methoden

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.

Referenties

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

Toepassingsgegevens

IHC

Gevalideerd door Selleck

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