We dissolved 1mg Aβ25–35 (A4559, Sigma) dry powder in 500 ul of 0.9% normal saline to prepare a 2 ug/ul solution and incubated for 7 days in a 37°C incubator after sonication, and the incubated Aβ25–35 oligomers were flocculent and stored in a refrigerator at 4°C for later use. Sodium pentobarbital was selected as an anaesthetic for rats, and we used a brain stereotaxic apparatus to fix and mark the coordinates of rats' heads. We prepared the head skin of the rats and drilled two holes (coordinates: 3 mm below the bregma and 2 mm on both sides of the midline) in the skull with a dental drill. We absorbed 5 ul of Aβ25–35 solution with a microsyringe and fixed it on the injection frame of the brain stereotaxic apparatus. We manipulated the instrument to lower the injection needle and probe into the hole, pierced the subdural by 2.6 mm, slowly injected the solution within 5 minutes, stopped for 15 minutes after the injection, and slowly withdrew the needle within 5 minutes. The skull hole was closed with paraffin, and the injection procedure was the same on the other side. After 7 days of injection, surviving rats can become AD models [19 (link), 20 (link)].
A4559
Manufactured by Merck Group
Sourced in United States
A4559 is a laboratory equipment product designed for general laboratory use. It serves as a tool for performing various tasks and experiments within a controlled laboratory environment. The core function of this product is to provide a standardized and reliable platform for conducting scientific investigations. No further details or interpretations are provided.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using a4559
1
Alzheimer's Disease Rat Model via Amyloid-Beta Injection
2
Astrocyte Activation Protocols
3
Inducing Alzheimer's-like Pathology in Rats
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Aβ peptide 25–35 (A4559; Sigma–Aldrich) was dissolved in saline solution (i.e., vehicle) at a concentration of 100 μM and incubated at 37°C for 4 days to induce Aβ 25–35 aggregation initiation. A total volume of 2.0 μl of Aβ protein was injected into each hippocampus based on the Paxinos and Watson brain atlas coordinates (anterior-posterior = −4.2 mm; lateral–lateral, ± 3.0 mm; ventral-medial, −3.0 mm) by stereotaxic microinjection using a Hamilton syringe and an infusion pump. After surgery, rats were returned to their home cages and were monitored for 10 days.
4
Preparation and Application of Aβ Oligomers
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Aβ oligomers were prepared as previously described [4 (link)]. Briefly, synthetic Aβ25−35 peptide (Sigma-Aldrich, A4559) was diluted in dimethyl sulfoxide (Sigma-Aldrich, D2650) and lyophilised overnight. To form Aβ25–35 oligomers, the lyophilised stock was resuspended in culture medium to the desired concentration and incubated at 37 °C for 24 h before the experiments. BECs were treated with Aβ25–35 at 10 μM for 24 h after transduced by lentivirus for 48 h. At least three independent experiments were performed.
5
Crocin Alleviates Alzheimer's Pathology
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Thirty mice were randomized into the sham group, Aβ25‐35 group, and Aβ25‐35+Crocin group (n = 10). Aβ25‐35 (A4559, Sigma‐Aldrich, USA) of 1 mL was dissolved in sterile saline (the final concentration of Aβ25‐35 solution was 5 µg/µL). The prepared Aβ25‐35 solution was incubated at 37°C for 7 days to form aggregates. An AD mouse model was established as previously described (Zhang et al., 2021 (link)). All mice were anesthetized with 3% isoflurane (792632, Sigma‐Aldrich, USA), and the brain was fixed using the brain stereotactic apparatus (7100‐R, RWD, China). Aβ25‐35 (2 µL) was injected into the hippocampus of mice, specifically, the anteroposterior (AP) was 2.0 mm, the mediolateral (ML) was 2.0 mm, and the dorsoventral (DV) was 1.7 mm. The mice in the sham group were injected with an equivalent volume of sterile saline in the same manner. After 24 h, the mice in the Aβ25‐35+Crocin group were injected intraperitoneally with 40 mg/kg of Crocin (purity ≥ 98%, 42553‐65‐1, Shanghai YuanYe Biotechnology Co., Ltd, dissolved in normal saline) daily for 14 consecutive days (Salem et al., 2022 (link); Wang et al., 2024 (link)). The other two groups of mice received the same amount of normal saline.
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