amyloid beta 42 human elisa kit  (Thermo Fisher)


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    Amyloid beta 42 Human ELISA Kit
    Description:
    The Human Beta Amyloid 1 42 Aβ42 ELISA research use only kit is to be used for the quantitative determination of human Aβ42 in samples e g cell culture supernatants tissue homogenates cerebrospinal fluid CSF etc using 96 well plates and a microplate reader The assay recognizes both natural and synthetic forms of human Aβ42 The anti human Aβ42 antibody used in this kit is capable of selectively detecting Aβ42 and not Aβ40 Aβ43 Performance characteristics • Sensitivity 10 pg mL • Standard curve range 15 6 1 000 pg mL • Sample types cell culture supernatants tissue homogenates cerebrospinal fluid CSF • Species cross reactivity human • Sample volume 50 μL prediluted • Total assay incubation time 4 hrs Principle of the method The Human Aβ42 kit is a solid phase sandwich enzyme linked immunosorbent assay ELISA A monoclonal antibody specific for the NH2 terminus of human Aβ has been coated onto the wells of the microtiter strips provided During the first incubation standards of known human Aβ42 content controls and unknown samples are pipetted into the wells and co incubated with a rabbit antibody specific for the COOH terminus of the 1 42 Aβ sequence This COOH terminal sequence is created upon cleavage of the analyzed precursor After washing bound rabbit antibody is detected by the addition of a horseradish peroxidase labeled anti rabbit antibody After a second incubation and washing to remove all of the unbound enzyme a substrate solution is added which is acted upon by the bound enzyme to produce color The intensity of this colored product is directly proportional to the concentration of human Aβ42 present in the original specimen Background information Beta amyloid peptide Aβ is a 40 to 43 amino acid peptide cleaved from amyloid precursor protein by β secretase e g BACE and a putative γ gamma secretase Increased release of the longer forms of Aβ peptide Aβ42 or Aβ43 which have a greater tendency to aggregate than Aβ40 occurs in individuals expressing certain genetic mutations expressing certain ApoE alleles or may involve other still undiscovered factors Many researchers theorize that it is this increased release of Aβ42 Aβ43 which leads to the abnormal deposition of Aβ Each Human Aβ42 ELISA kit is validated for sensitivity specificity precision and lot to lot consistency See product insert for more information on validation Related links Learn more about ELISA kits Learn more about other immunoassays Please refer to the ELISA protocol for specific reference citations
    Catalog Number:
    khb3441
    Price:
    None
    Category:
    Kits and Assays
    Applications:
    Protein Assays and Analysis|Protein Biology
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    Structured Review

    Thermo Fisher amyloid beta 42 human elisa kit
    The Human Beta Amyloid 1 42 Aβ42 ELISA research use only kit is to be used for the quantitative determination of human Aβ42 in samples e g cell culture supernatants tissue homogenates cerebrospinal fluid CSF etc using 96 well plates and a microplate reader The assay recognizes both natural and synthetic forms of human Aβ42 The anti human Aβ42 antibody used in this kit is capable of selectively detecting Aβ42 and not Aβ40 Aβ43 Performance characteristics • Sensitivity 10 pg mL • Standard curve range 15 6 1 000 pg mL • Sample types cell culture supernatants tissue homogenates cerebrospinal fluid CSF • Species cross reactivity human • Sample volume 50 μL prediluted • Total assay incubation time 4 hrs Principle of the method The Human Aβ42 kit is a solid phase sandwich enzyme linked immunosorbent assay ELISA A monoclonal antibody specific for the NH2 terminus of human Aβ has been coated onto the wells of the microtiter strips provided During the first incubation standards of known human Aβ42 content controls and unknown samples are pipetted into the wells and co incubated with a rabbit antibody specific for the COOH terminus of the 1 42 Aβ sequence This COOH terminal sequence is created upon cleavage of the analyzed precursor After washing bound rabbit antibody is detected by the addition of a horseradish peroxidase labeled anti rabbit antibody After a second incubation and washing to remove all of the unbound enzyme a substrate solution is added which is acted upon by the bound enzyme to produce color The intensity of this colored product is directly proportional to the concentration of human Aβ42 present in the original specimen Background information Beta amyloid peptide Aβ is a 40 to 43 amino acid peptide cleaved from amyloid precursor protein by β secretase e g BACE and a putative γ gamma secretase Increased release of the longer forms of Aβ peptide Aβ42 or Aβ43 which have a greater tendency to aggregate than Aβ40 occurs in individuals expressing certain genetic mutations expressing certain ApoE alleles or may involve other still undiscovered factors Many researchers theorize that it is this increased release of Aβ42 Aβ43 which leads to the abnormal deposition of Aβ Each Human Aβ42 ELISA kit is validated for sensitivity specificity precision and lot to lot consistency See product insert for more information on validation Related links Learn more about ELISA kits Learn more about other immunoassays Please refer to the ELISA protocol for specific reference citations
    https://www.bioz.com/result/amyloid beta 42 human elisa kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    amyloid beta 42 human elisa kit - by Bioz Stars, 2021-03
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    Enzyme-linked Immunosorbent Assay:

    Article Title: Evaluation of Plant Phenolic Metabolites as a Source of Alzheimer's Drug Leads
    Article Snippet: .. They were frozen until assay of amyloid beta 42 levels with ELISA (Invitrogen, USA) [ ]. .. ELISA and Measurement of Amyloid Beta 42 Samples were centrifuged at 16,000 g for 20 min at 4°C.

    Article Title: A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer's disease
    Article Snippet: To measure Aβ levels in conditioned media of APP-HEK293 cells, media of 72-h cell culture was collected, mixed with 4X sample buffer (50 mM Tris-HCl pH6.8, 2% SDS, 10% glycerol, 1% β-mercaptoethanol, 12.5 mM EDTA, 0.02% bromophenol blue), and boiled at 95°C for 10 min. One μl of each sample was spotted on nitrocellulose membrane for dot blot analysis. .. ELISA GuHCl extracted brain samples prepared in the same way as dot blot assays were diluted 1:1000, and ELISA analyses of Aβ42 were performed according to manufacturer’s instructions (Thermo Fisher Scientific, KHB3441). .. Immunofluorescence microscopy Paraformaldehyde-fixed brain tissues were sectioned at 30 μm thickness.

    Article Title: Fingolimod modulates multiple neuroinflammatory markers in a mouse model of Alzheimer’s disease
    Article Snippet: ELISA assay Dissected frontal brain tissue was homogenized with 8 × 5 M guanidine HCl buffer to analyze the level of total (soluble + insoluble) Aβ. .. To determine Aβ levels, human Aβ40 and Aβ42 ELISA kits (Invitrogen, Grand Island, NY) were used according to manufacturer’s specifications. ..

    Article Title: Suppression of glymphatic fluid transport in a mouse model of Alzheimer’s disease
    Article Snippet: We used 125 I-Aβ40 without added aprotinin, a potential inhibitor of LRP1-mediated transport. .. ELISA kits for Aβ40 (KHB 3481) and Aβ42 (KHB 3441) were obtained from Invitrogen (Camarilla, CA, USA) and Aβ oligomer ELISA kit (BEK-2215-1P) from Biosensis (Termecula, CA, USA). .. Mice were anesthetized as indicated above, fixed to a stereotactic frame, cisterna magna exposed and cannulated with a 30G needle ( ; ).

    Article Title: Protective effects of 7,8-dihydroxyflavone on neuropathological and neurochemical changes in a mouse model of Alzheimer’s disease
    Article Snippet: .. To determine Aβ levels , human Aβ40 and Aβ42 ELISA kits (Invitrogen, Grand Island, NY) were used according to manufacturer’s specifications. ..

    Article Title: Suppression of glymphatic fluid transport in a mouse model of Alzheimer’s disease
    Article Snippet: The supernatant was used to determine levels of soluble Aβ oligomers. .. Levels (pmol/g brain tissue) of human Aβ40, Aβ42 and soluble Aβ oligomers were determined using ELISA kits [Aβ40 (KHB 3481) and Aβ42 (KHB 3441); Invitrogen (Camarilla, CA, USA] and following the manufacturers’ instructions. .. Since soluble Aβ oligomers vary in size and number of oligomers the unit is arbitrary and is indicated as equivalent to Aβ.

    Article Title: Disease-Modifying Effects of M1 Muscarinic Acetylcholine Receptor Activation in an Alzheimer’s Disease Mouse Model
    Article Snippet: Total A β 40 and A β 42 immunoreactive surface area was then measured in a blinded manner using MetaMorph 5.0 software (Molecular Devices). .. hA β 40 and hA β 42 levels in conditioned media from primary neuronal cultures as well as soluble and insoluble A β 40 and A β 42 tissue homogenates from biochemically fractionated mouse brains were measured using human A β 40 and A β 42 ELISA kits according to the manufacturer’s protocols (Biosource, Invitrogen). .. Insoluble amyloid fractions from mouse tissue homogenates containing 70% formic acid were first neutralized by performing a 1:100 dilution in a solution of 1.0 M Tris (pH 11) prior to performing dilution series in ELISA diluent buffer supplied with ELISA kits.

    Dot Blot:

    Article Title: A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer's disease
    Article Snippet: To measure Aβ levels in conditioned media of APP-HEK293 cells, media of 72-h cell culture was collected, mixed with 4X sample buffer (50 mM Tris-HCl pH6.8, 2% SDS, 10% glycerol, 1% β-mercaptoethanol, 12.5 mM EDTA, 0.02% bromophenol blue), and boiled at 95°C for 10 min. One μl of each sample was spotted on nitrocellulose membrane for dot blot analysis. .. ELISA GuHCl extracted brain samples prepared in the same way as dot blot assays were diluted 1:1000, and ELISA analyses of Aβ42 were performed according to manufacturer’s instructions (Thermo Fisher Scientific, KHB3441). .. Immunofluorescence microscopy Paraformaldehyde-fixed brain tissues were sectioned at 30 μm thickness.

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    Thermo Fisher aβ42
    An autophagy-inducing compound ML246 reduces amyloid load in an autophagy-dependent manner in vitro and in vivo. (A) Chemical structure of ML246. (B) Dot-blot assays (left) and quantification (right) of secreted <t>Aβ42</t> levels in conditioned media of HEK293 cells stably expressing APP treated with vehicle (DMSO) or ML246 for 24 h, immunostained with anti-Aβ42 antibody. Cells were transfected with non-targeting control (NC) or ATG7 siRNA 24 h prior to ML246 treatment. Results are quantified from 4 independent experiments. (C) Representative images (left) and quantification (right) of TUNEL signals (red) in WT primary cortical neurons treated with conditioned media from ( B ) for 24 h. Nuclei were stained with DAPI. Scale bar, 100 μm. N = 10 fields (each field containing 20–30 neurons). (D, E) Representative images (upper) and quantification (lower) of dot-blot assays on soluble (D) and insoluble (E) Aβ42 levels in brain samples of 6-month old 5XFAD and 5XFAD Becn1 +/- KO mice after 5 weeks of ML246 treatment, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 4–5 mice in each group were shown. Results represent mean ± s.e.m. NS, not significant; *, P
    Aβ42, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aβ42/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
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    aβ42 - by Bioz Stars, 2021-03
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    An autophagy-inducing compound ML246 reduces amyloid load in an autophagy-dependent manner in vitro and in vivo. (A) Chemical structure of ML246. (B) Dot-blot assays (left) and quantification (right) of secreted Aβ42 levels in conditioned media of HEK293 cells stably expressing APP treated with vehicle (DMSO) or ML246 for 24 h, immunostained with anti-Aβ42 antibody. Cells were transfected with non-targeting control (NC) or ATG7 siRNA 24 h prior to ML246 treatment. Results are quantified from 4 independent experiments. (C) Representative images (left) and quantification (right) of TUNEL signals (red) in WT primary cortical neurons treated with conditioned media from ( B ) for 24 h. Nuclei were stained with DAPI. Scale bar, 100 μm. N = 10 fields (each field containing 20–30 neurons). (D, E) Representative images (upper) and quantification (lower) of dot-blot assays on soluble (D) and insoluble (E) Aβ42 levels in brain samples of 6-month old 5XFAD and 5XFAD Becn1 +/- KO mice after 5 weeks of ML246 treatment, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 4–5 mice in each group were shown. Results represent mean ± s.e.m. NS, not significant; *, P

    Journal: PLoS Genetics

    Article Title: A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer's disease

    doi: 10.1371/journal.pgen.1006962

    Figure Lengend Snippet: An autophagy-inducing compound ML246 reduces amyloid load in an autophagy-dependent manner in vitro and in vivo. (A) Chemical structure of ML246. (B) Dot-blot assays (left) and quantification (right) of secreted Aβ42 levels in conditioned media of HEK293 cells stably expressing APP treated with vehicle (DMSO) or ML246 for 24 h, immunostained with anti-Aβ42 antibody. Cells were transfected with non-targeting control (NC) or ATG7 siRNA 24 h prior to ML246 treatment. Results are quantified from 4 independent experiments. (C) Representative images (left) and quantification (right) of TUNEL signals (red) in WT primary cortical neurons treated with conditioned media from ( B ) for 24 h. Nuclei were stained with DAPI. Scale bar, 100 μm. N = 10 fields (each field containing 20–30 neurons). (D, E) Representative images (upper) and quantification (lower) of dot-blot assays on soluble (D) and insoluble (E) Aβ42 levels in brain samples of 6-month old 5XFAD and 5XFAD Becn1 +/- KO mice after 5 weeks of ML246 treatment, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 4–5 mice in each group were shown. Results represent mean ± s.e.m. NS, not significant; *, P

    Article Snippet: ELISA GuHCl extracted brain samples prepared in the same way as dot blot assays were diluted 1:1000, and ELISA analyses of Aβ42 were performed according to manufacturer’s instructions (Thermo Fisher Scientific, KHB3441).

    Techniques: In Vitro, In Vivo, Dot Blot, Stable Transfection, Expressing, Transfection, TUNEL Assay, Staining, Mouse Assay, Labeling

    Autophagosomal sequestration of Aβ42 in brain of autophagy-hyperactive mice. (Left) Scheme of immunoisolation of autophagosomes from brain of 12-week old 5XFAD Becn1 FA/FA mice expressing GFP-LC3. Briefly, post-nucleus extracts of the brain lysates was obtained by centrifugation at a low speed of 1,000 xg. Autophagosomes were enriched by centrifugation at a high speed of 20,000 xg, and pulled down by an anti-GFP antibody using magnetic beads. (Right) Western blot detection of Aβ42 fibrillar and oligomeric species inside autophagosomes immunoprecipitated by GFP antibody as in the scheme. A known autophagy cargo p62 serves as a positive control, and a cytosolic enzyme GAPDH is a negative control.

    Journal: PLoS Genetics

    Article Title: A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer's disease

    doi: 10.1371/journal.pgen.1006962

    Figure Lengend Snippet: Autophagosomal sequestration of Aβ42 in brain of autophagy-hyperactive mice. (Left) Scheme of immunoisolation of autophagosomes from brain of 12-week old 5XFAD Becn1 FA/FA mice expressing GFP-LC3. Briefly, post-nucleus extracts of the brain lysates was obtained by centrifugation at a low speed of 1,000 xg. Autophagosomes were enriched by centrifugation at a high speed of 20,000 xg, and pulled down by an anti-GFP antibody using magnetic beads. (Right) Western blot detection of Aβ42 fibrillar and oligomeric species inside autophagosomes immunoprecipitated by GFP antibody as in the scheme. A known autophagy cargo p62 serves as a positive control, and a cytosolic enzyme GAPDH is a negative control.

    Article Snippet: ELISA GuHCl extracted brain samples prepared in the same way as dot blot assays were diluted 1:1000, and ELISA analyses of Aβ42 were performed according to manufacturer’s instructions (Thermo Fisher Scientific, KHB3441).

    Techniques: Mouse Assay, Expressing, Centrifugation, Magnetic Beads, Western Blot, Immunoprecipitation, Positive Control, Negative Control

    ML246 and voluntary exercise decrease cerebral amyloid plaques and ameliorate memory deficits in 5XFAD AD mice. (A, B) Representative images (upper) and quantification (lower) of dot-blot assays on soluble (A) and insoluble (B) Aβ42 levels in brain samples of 6-month old 5XFAD and 5XFAD Becn1 +/- KO mice after 4 months of voluntary running, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 4–5 mice in each group were shown. (C) Representative images (left) and quantification (right) of amyloid deposits stained by Thioflavin S in brain of 6-month old 5XFAD mice, and 5XFAD mice subject to 5 weeks of ML246 treatment or 4 months of voluntary exercise. Scale bar: 500 μm. Results represent mean ± s.e.m. N = 6–8. *, P

    Journal: PLoS Genetics

    Article Title: A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer's disease

    doi: 10.1371/journal.pgen.1006962

    Figure Lengend Snippet: ML246 and voluntary exercise decrease cerebral amyloid plaques and ameliorate memory deficits in 5XFAD AD mice. (A, B) Representative images (upper) and quantification (lower) of dot-blot assays on soluble (A) and insoluble (B) Aβ42 levels in brain samples of 6-month old 5XFAD and 5XFAD Becn1 +/- KO mice after 4 months of voluntary running, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 4–5 mice in each group were shown. (C) Representative images (left) and quantification (right) of amyloid deposits stained by Thioflavin S in brain of 6-month old 5XFAD mice, and 5XFAD mice subject to 5 weeks of ML246 treatment or 4 months of voluntary exercise. Scale bar: 500 μm. Results represent mean ± s.e.m. N = 6–8. *, P

    Article Snippet: ELISA GuHCl extracted brain samples prepared in the same way as dot blot assays were diluted 1:1000, and ELISA analyses of Aβ42 were performed according to manufacturer’s instructions (Thermo Fisher Scientific, KHB3441).

    Techniques: Mouse Assay, Dot Blot, Labeling, Staining

    Becn1 FA/FA mutation ameliorates cerebral Aβ accumulation, memory deficits and mortality of Alzheimer’s mouse models. (A-B) Dot-blot assays and quantification of soluble (A) and insoluble (B) Aβ42 levels in homogenated brain samples of 6-month old 5XFAD Becn1 +/+ and 5XFAD Becn1 FA/FA mice, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 6 mice in each group were shown. ***, P

    Journal: PLoS Genetics

    Article Title: A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer's disease

    doi: 10.1371/journal.pgen.1006962

    Figure Lengend Snippet: Becn1 FA/FA mutation ameliorates cerebral Aβ accumulation, memory deficits and mortality of Alzheimer’s mouse models. (A-B) Dot-blot assays and quantification of soluble (A) and insoluble (B) Aβ42 levels in homogenated brain samples of 6-month old 5XFAD Becn1 +/+ and 5XFAD Becn1 FA/FA mice, immunostained with anti-Aβ42 antibody. Total protein loading was labeled by Ponceau S. Triplicate experiments from 6 mice in each group were shown. ***, P

    Article Snippet: ELISA GuHCl extracted brain samples prepared in the same way as dot blot assays were diluted 1:1000, and ELISA analyses of Aβ42 were performed according to manufacturer’s instructions (Thermo Fisher Scientific, KHB3441).

    Techniques: Mutagenesis, Dot Blot, Mouse Assay, Labeling