Proteins were resolved on 8%, 10%, 15%, or 4-20% tris-glycine gels, and for amyloid-β or Ub immunoblotting 16% tris-tricine gel was used62 (link), transferred to nitrocellulose membranes (GE Whatman, 0.2 µm Pore Size). Western blot images were visualized by enhanced chemiluminescence (ECL). The images were captured by using an ImageQuant™ LAS 4000 imaging system (GE Healthcare) and quantitated by Image Studio™ Lite (LI-COR Biosciences). Primary antibodies were used at the following dilutions: 6E10 anti-amyloid-β (1:1000, Biolegend, Cat. no. 803015); anti-PSEN1 (1:1,000, Cell Signaling Technology, Cat. no. 5643); anti-β-tubulin (1:1,000, Abcam, Cat. no. AB24629); anti-UBB+1 (1:1000, custom made by Sigma for our lab), anti-ubiquitin (1:2000, DAKO, Cat. no. Z0458), anti-UCH-L1 (1:1000, Abcam, Cat. no. ab8189), anti-GAPDH (1:10,000, Sigma, Cat. no. G9545), anti-β-actin (1:10,000, Santa Cruz, Cat. no. sc-47778).
6e10 anti amyloid β
Manufactured by BioLegend
Sourced in United States
6E10 is a monoclonal antibody that binds to amino acids 1-16 of the amyloid-β peptide. It can be used in various research applications involving the detection and study of amyloid-β.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Merck Group (1 mentions)
Image studio lite, by LI COR (1 mentions)
Anti β tubulin, by Abcam (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Imagequant las 4000 imaging system, by GE Healthcare (1 mentions)
Anti uch l1, by Abcam (1 mentions)
Anti ubiquitin, by Agilent Technologies (1 mentions)
Whatman, by Cytiva (1 mentions)
Las 4000, by Cytiva (1 mentions)
Glial fibrillary acidic protein (gfap), by Abcam (1 mentions)
Deltavision system, by GE Healthcare (1 mentions)
Confocal laser scanning microscope, by Zeiss (1 mentions)
P tau thr181, by Cell Signaling Technology (1 mentions)
4 protocols using 6e10 anti amyloid β
1
Western Blot Analysis of Amyloid-β and Ubiquitin
2
Dot Blot Immunodetection of Alzheimer's Proteins
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Dot blot was performed as previously described29 (link). Primary antibodies were used at the following dilutions: 6E10 anti-amyloid-β (1:1000, Biolegend, Cat. no. 803015); anti-UBB+1 (1:1000, custom made by Sigma for our lab), anti-MC1 (1:1000, kindly donated by P. Davies) and anti-PHF-1 (1:1000, kindly donated by P. Davies).
3
Multimodal Imaging of Amyloid, Lactate, and Glia
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Paraffin sections of brain tissue were dewaxed in different concentrations of organic reagents, and antigen retrieval was performed with ethylenediaminetetraacetic acid in a microwave oven. Tissue slices were incubated with an autofluorescence quencher for 5 min to eliminate autofluorescence. The primary antibody (anti-β-amyloid (6E10) (BioLegend), MCT4 (Santa Cruz Biotechnology) and GFAP (Abcam)) diluted in 5% BSA was added to cover the tissue after the slices were dried. Then, the slices were placed in the wet box and incubated overnight at 4 °C, and the immunoreaction was detected using the fluorescence-conjugated secondary antibody. The nuclei were stained with DAPI, and the tissue sections were sealed with an anti-fluorescence quencher. Images were captured using a confocal laser scanning microscope (Zeiss, Jena, Germany).
For thioflavin S (ThS) staining, after incubation with β-amyloid (6E10) antibody and the fluorescent secondary antibody, the tissue sections were stained for 8 min with a solution of 0.3% ThS in 50% ethanol. Images captured using a DeltaVision system (GE Healthcare, Little Chalfont, UK).
For thioflavin S (ThS) staining, after incubation with β-amyloid (6E10) antibody and the fluorescent secondary antibody, the tissue sections were stained for 8 min with a solution of 0.3% ThS in 50% ethanol. Images captured using a DeltaVision system (GE Healthcare, Little Chalfont, UK).
4
Quantitative Analysis of Alzheimer's Markers
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Mouse brains were processed into 5 μm thick slices with paraffin embedding and dewaxed in a gradient concentration organic solvent. Tissue sections were placed in a microwave oven for antigen retrieval, immersed in 3% hydrogen peroxide solution at room temperature and protected from light for 25 min to block endogenous peroxidase. After the slices were dried, the primary antibody (anti-β-amyloid (6E10) (BioLegend, San Diego, USA), p-tau (Thr 181) (Cell Signaling Technology, Danvers, USA), MCT2 (ABclonal, Wuhan, China) and MCT4 (Santa Cruz Biotechnology, Santa Cruz, USA)) diluted in 5% bovine serum albumin (BSA) was added to cover the tissue. The sections were placed in a wet box and incubated overnight at 4 °C, and the HRP-conjugated secondary antibody and diaminobenzidine tetrahydrochloride were used. The nuclei were stained with hematoxylin, and the tissue sections were sealed with neutral gum. Images were captured using a compound microscope (COIC, Chongqing, China). The analysis of protein level was performed in ImageJ's “analyze particles” function used for counting relative areas, and thresholds were consistent across all groups.
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