The following antibodies were used in this study: antibodies against β-amyloid (MOAB-2) were purchased from Novus Biologicals (Littleton, CO, USA). Antibodies against α-synuclein (phospho S129) (ab51253), Iba1 (ab178846), and NeuN (ab177487) were purchased from Abcam (CA, UK). Antibodies against Tau-5 (MA5-12808), GFAP (PA5-16291), and PGRN (40-3400) were purchased from Thermo Fisher Scientific (Bridgewater, NJ, USA). Antibody against TDP-43 (10782-2-AP) was purchased from Proteintech (IL, USA). Antibodies against LC3B (2775S) and GAPDH (2118) were purchased from Cell Signaling Technology (Danvers, MA, USA). Fluorescence-labeled secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA). DAPI (H-1200) was purchased from VECTOR Laboratories (Burlingame, CA, USA). RNeasy Mini Kit (74104) was purchased from Qiagen (MD, USA). High-Capacity cDNA Reverse Transcription Kit (43-688-14) was purchased from Applied Biosystems™ (Waltham, MA, USA). SYBR™ Green PCR Master Mix (4309155) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Moab 2
Manufactured by Novus Biologicals
Sourced in United States
MOAB-2 is a high-performance liquid chromatography (HPLC) system designed for efficient and accurate separation and analysis of a wide range of biological samples. The core function of MOAB-2 is to provide precise and reliable chromatographic separation, enabling researchers to isolate and quantify target analytes with confidence.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Fluorescence labeled secondary antibody, by Jackson ImmunoResearch (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Dapi h 1200, by Vector Laboratories (1 mentions)
Hrp conjugated goat anti mouse igg, by Novus Biologicals (1 mentions)
Hrp conjugated goat anti mouse igg, by Boster Bio (1 mentions)
Nlrp3, by Novus Biologicals (1 mentions)
Gsdmd, by Abcam (1 mentions)
Cytation 5, by Agilent Technologies (1 mentions)
4 protocols using moab 2
1
Antibody Resources for Neurodegenerative Research
2
Immunostaining Protocol for IgG and Aβ
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Immunostaining for IgG was carried out as previously described [26 (link), 27 (link)]. Briefly, the 30 μm-thick sections were washed in 0.01 M PBS three times in order to wash out cryoprotectant and then incubated in 3% hydrogen peroxide (methanol: 30% H2O2 = 1:1 in 0.01 M PBS) for 20 min, permeabilized with PBS containing 0.25%Triton X-100 (PBST, 3 × 10 min). The sections were blocked in 5% normal goat serum at RT for 1 h, followed incubation with HRP-conjugated goat anti-mouse IgG (Boster Biological Technology, Wuhan, China) overnight at 4 °C. Free-floating sections were subsequently washed in PBS three times. Sections were incubated in 0.05% diaminobenzidine (DAB) to visualize reaction products and then washed with PBS (3 × 5 min), mounted to a glass slide. Then sample was dried in air overnight, dehydrated using graded ethanol, cleared in xylene before cover-slipped with Neutral balsam. Images were captured under a light microscope (Olympus Corporation, Tokyo, Japan).
Immunostaining for Aβ was followed with the above procedures, incubated with primary antibody MOAB2 (Novus) followed by HRP-conjugated goat anti-mouse IgG secondary antibody.
Immunostaining for Aβ was followed with the above procedures, incubated with primary antibody MOAB2 (Novus) followed by HRP-conjugated goat anti-mouse IgG secondary antibody.
3
Immunohistochemical Analysis of Hippocampal Slices
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The hippocampal slices were prepared at a thickness of 4 μm and then incubated with 0.3% hydrogen peroxide (which was added dropwise) at room temperature for 10 minutes to block endogenous peroxidase activity. The slices were washed 3 times with phosphate-buffered saline (PBS) and blocked with 5% bovine serum albumin for 30 minutes. The slices were incubated overnight with Aβ (MOAB-2, recognizes unaggregated, oligomeric and fibrillar forms of beta amyloid 42 and unaggregated beta amyloid 40, 1:500, Novus, USA), NLRP3 (1:50, Novus, USA), and GSDMD (1:1000, Abcam, UK) antibodies. Subsequently, the slices were washed 3 times with PBS and incubated with biotin secondary antibodies for 20 minutes and then treated with 3,3′-diaminobenzidine for 3 to 5 minutes. After washing, they were counterstained with haematoxylin and mounted after dehydration [34 (link)]. The slices were observed using a Cytation 5 (BioTek, USA) image reader. ImageJ software was used to analyse the average optical density (AOD) values and positive cell counts.
4
Molecular Profiling of Alzheimer's Pathology
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Protein expression related to amyloid-β accumulation (APP, BACE-1, presenilin, amyloid-β), neuronal regeneration (NGF, BDNF), and inflammation (COX-2) was investigated using Western blot analysis. A bicinchoninic acid (BCA) protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to assess protein concentration, followed by Western blot analysis. Protein (15 μg) extracted from brain tissue was separated using 10% sodium dodecyl sulfate (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was blocked in a buffer containing antibodies of APP (amyloid precursor protein, 1:1000, 2452 S, Cell Signaling, Danvers, MA, USA), BACE-1 (β-secretase 1, 1:1000, 5606 S, Cell Signaling, USA), presenilin (1:1000, 5643 S, Cell Signaling, USA), amyloid-β (1:1000, MOAB-2, NOVUS Biological, USA), NGF (nerve growth factor, 1:1000, PA5-29425, Invitrogen, Waltham, MA, USA), BDNF (brain-derived neurotrophic factor; 1:1000, 47,808 S, Cell Signaling, USA), and COX-2 (cycloxygenase-2, 1:1000, 12,282 S, Cell Signaling, USA) at 4 °C overnight. The membrane was incubated with specific secondary antibodies (1:2000) at room temperature for 1 h, then further incubated with enhanced chemiluminescence (ECL) substrate and the bands were detected using a Chemi-Doc XRS system. Protein loading was evaluated using anti-β-actin antibody.
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