Khb3441

Manufactured by Thermo Fisher Scientific

Sourced in United States

The KHB3441 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed to perform a core function within a laboratory setting. However, a detailed and unbiased description of the product's specific capabilities and intended use cannot be provided without the risk of extrapolation or interpretation. As a result, a comprehensive factual description of the KHB3441 is not available.

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Lab products found in correlation

Khb3481, by Thermo Fisher Scientific (30 mentions) Versamax tunable microplate reader, by Molecular Devices (4 mentions) Kmb3441, by Thermo Fisher Scientific (2 mentions) Kmb3481, by Thermo Fisher Scientific (2 mentions) Versamax, by Molecular Devices (2 mentions) Spectramax i3x multi mode microplate reader, by Molecular Devices (1 mentions) Ripa cell lysis buffer, by Thermo Fisher Scientific (1 mentions) Ripa buffer, by Merck Group (1 mentions) Phosphatase inhibitor cocktail, by AG Scientific (1 mentions) Kho0631, by Thermo Fisher Scientific (1 mentions) Tr15421, by Thermo Fisher Scientific (1 mentions) Mak043, by Merck Group (1 mentions) Ab87546, by Abcam (1 mentions) Coomassie bradford protein assay, by Thermo Fisher Scientific (1 mentions) Mpm 6, by Bio-Rad (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Spectramax m2, by Molecular Devices (1 mentions) 8 isoprostane affinity sorbent, by Cayman Chemical (1 mentions) Eicosanoid column affinity buffer, by Cayman Chemical (1 mentions) Elisa technical guide, by Thermo Fisher Scientific (1 mentions)

53 protocols using khb3441

Hippocampal Protein Extraction and Analysis

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The 6 right hippocampi were weighed in each group and homogenized with 8 times the volume of mixed liquor of 5 M guanidine hydrochloride, 50 mM Tris hydrochloric acid (pH 8.0), and 1 mM PMSF on ice. Then the hippocampi were centrifuged with 16000 r/min, at 4°C, for 20 min, and the supernatant was obtained. The diluted samples were prepared by separately mixing the supernatant with a standard diluent of 3200 times the volume (KHB3441, Invitrogen, USA) and 800 times the volume (KHB3441, Invitrogen, USA) with the hippocampus of the model and EA groups. The samples in the normal control group were not diluted.

Wang X., Miao Y., Abulizi J., Li F., Mo Y., Xue W, & Li Z. (2016). Improvement of Electroacupuncture on APP/PS1 Transgenic Mice in Spatial Learning and Memory Probably due to Expression of Aβ and LRP1 in Hippocampus. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 7603975.

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Quantification of Amyloid-Beta Peptides

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N2a-wt and N2a-APPwt cells were plated and treated as described in the “Cell Treatment” section. After 24 h the culture medium was replaced with serum-free medium for another 24 h. The supernatant was further collected and centrifuged at 16,000× g for 10 min at 4 °C. The levels of Aβ40 and Aβ42 were measured using a sandwich ELISA kit (KHB3481 and KHB3441 from Invitrogen) following the manufacturer’s instructions. The results are expressed in pg/mL.

Silva A., Pereira M., Carrascal M.A., Brites G., Neves B., Moreira P., Resende R., Silva M.M., Santos A.E., Pereira C, & Cruz M.T. (2020). Calcium Modulation, Anti-Oxidant and Anti-Inflammatory Effect of Skin Allergens Targeting the Nrf2 Signaling Pathway in Alzheimer’s Disease Cellular Models. International Journal of Molecular Sciences, 21(20), 7791.

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Plasma Aβ and FI Quantification

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Plasma concentrations of Aβ were determined by ELISA, which either detects both Aβ40 and Aβ42 [53 (link)] or specifically measures each peptide. In the latter case, ELISA kits from Invitrogen (KHB3481 and KHB3441) were used. Plasma concentration of FI was also measured by ELISA using a kit from GenWay Biotech. The commercial kits were used by following the manufacturers’ instructions.

Yang L., Bhattacharya A., Li Y, & Zhang Y. (2017). Anticoagulants inhibit proteolytic clearance of plasma amyloid beta. Oncotarget, 9(5), 5614-5626.

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Quantification of Amyloid-β Peptides

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Amyloid-β₄₀ (Invitrogen; KHB3481) and amyloid-β₄₂ (Invitrogen; KHB3441) ELISAs were conducted following the manufacturer’s guidelines. Amyloid-β₄₀ and amyloid-β₄₂ concentrations (pg/ml) were determined by fitting the standard curve. All samples and standards were run in triplicate.

Morrone C.D., Bazzigaluppi P., Beckett T.L., Hill M.E., Koletar M.M., Stefanovic B, & McLaurin J. (2019). Regional differences in Alzheimer’s disease pathology confound behavioural rescue after amyloid-β attenuation. Brain, 143(1), 359-373.

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Quantifying Cardiac Aβ Peptides

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Aβ40 and Aβ42 peptides levels were detected using a commercial ELISA kit (Invitrogen KHB3481 and KHB3441, respectively). Briefly, cardiac specimens for each mouse were collected at the end of the experimental study and homogenized with RIPA cell lysis buffer (Thermo Scientific), supplemented with phosphatases and protease inhibitors (Thermo Scientific, Waltham, MA). Then, ELISA was performed using an equal amount of total cardiac protein following the company’s specifications, and the plate was evaluated at 450 nm with a spectrophotometer reader (SpectraMax i3x Multi-mode Microplate Reader, Molecular Devices).

Elia A., Parodi-Rullan R., Vazquez-Torres R., Carey A., Javadov S, & Fossati S. (2023). Amyloid β induces cardiac dysfunction and neuro-signaling impairment in the heart of an Alzheimer’s disease model. bioRxiv.

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Quantification of Insoluble Amyloid-β

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The hippocampal and cortical homogenates were collected for supernatant fraction and allowed to precipitate. The pellet was further re-suspended in 70% formic acid, received sonication for 10 s, and was centrifuged at 17,000 rpm for 1 h at 4°C. The supernatant was collected for the insoluble Aβ ELISA assay. The insoluble levels of Aβ1-40 and Aβ1-42 were determined by using commercial kits (KHB3481 and KHB3441, Invitrogen, United States), according to the manufacturer’s instructions.

Tan Y., Wang X., Zhang J., Zhang H., Li H., Peng T., Chen W., Wei P., Liu Z., He F., Li J., Ding H., Li N., Wang Z., Zhang Z, & Hua Q. (2022). NeuroProtect, a Candidate Formula From Traditional Chinese Medicine, Attenuates Amyloid-β and Restores Synaptic Structures in APP/PS1 Transgenic Mice. Frontiers in Pharmacology, 13, 850175.

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Quantifying Amyloid-Beta Peptides

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The mice brains were homogenized in 8× mass of 5 M guanidine HCl/50 mM tris-HCl (pH 8.0) and incubated at room temperature for 3 hours. Then, the samples were diluted with cold reaction buffer (PBS with 5% BSA and 0.03% Tween 20, supplemented with protease inhibitor cocktail) and centrifuged at 16,000g for 20 min at 4°C. The supernatant was analyzed by human Aβ40 and Aβ42 ELISA kit according to the manufacturer’s instructions (KHB3481 and KHB3441, respectively, Invitrogen). The Aβ concentrations were determined by comparison with the standard curve.

Chen C., Ahn E.H., Kang S.S., Liu X., Alam A, & Ye K. (2020). Gut dysbiosis contributes to amyloid pathology, associated with C/EBPβ/AEP signaling activation in Alzheimer’s disease mouse model. Science Advances, 6(31), eaba0466.

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Amyloid-beta Protein Extraction and Quantification

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RIPA extraction (defined as soluble) and guanidine extraction (defined as insoluble) proteins were obtained from mice hippocampus and cortex. Cells supernatants were collected 48 hours post-transfection or 6 hours post-drug-treatment. Human and mouse Aβ1–40 and Aβ1–42 levels were measured by ELISA following the manufacture’s protocol (#KHB3481 and #KHB3441 for the human, #KMB3481 and #KMB3441 for the mouse, Invitrogen, Waltham, MA, USA).

Hernandez-Rapp J., Rainone S., Goupil C., Dorval V., Smith P.Y., Saint-Pierre M., Vallée M., Planel E., Droit A., Calon F., Cicchetti F, & Hébert S.S. (2016). microRNA-132/212 deficiency enhances Aβ production and senile plaque deposition in Alzheimer’s disease triple transgenic mice. Scientific Reports, 6, 30953.

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Quantifying Mouse Brain Amyloid-Beta 42

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ELISA kits for mouse Aβ42 were purchased from Invitrogen (KHB3441), and the assays were performed on lysates of mouse brains following the manufacturer’s instructions.

Li T., Yin L., Kang X., Xue W., Wang N., Zhang J., Yuan P., Lin L, & Li Y. (2022). TFEB acetylation promotes lysosome biogenesis and ameliorates Alzheimer’s disease–relevant phenotypes in mice. The Journal of Biological Chemistry, 298(12), 102649.

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Quantifying Soluble Amyloid-Beta Oligomers

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To measure the concentrations of soluble Aβ oligomers, each hemibrain sample was homogenized in 8-fold volumes of homogenization medium, as described above. To quantitate total levels of Aβ42, each hemibrain was extracted in 8-fold volumes of cold 5 M guanidine HCl plus 50 mM Tris HCl (pH 8.0) buffer and centrifuged at 20,000 g for 1 h at 4°C to remove insoluble material. Final guanidine HCl concentrations were below 0.1 M. Protein concentrations were determined by a BCA protein assay kit (Pierce). Supernatant fractions were analyzed by well-established human Aβ ELISA kits specific to oligomeric forms of Aβ (27725, IBL America, Minneapolis, MN, USA) and Aβ42 (KHB3441, Invitrogen) according to the protocols of the manufacturers. Optical densities at 450 nm of each well were read on a VersaMax tunable microplate reader (Molecular Devices, Sunnyvale, CA, USA), and sample Aβ oligomer and Aβ42 concentrations were determined by comparison with the respective standard curves. Aβ oligomer and Aβ42 concentration values were normalized to total brain protein concentrations and expressed in picograms and nanograms per milligram of total protein, respectively.

Devi L, & Ohno M. (2015). Effects of BACE1 Haploinsufficiency on APP Processing and Aβ Concentrations in Male and Female 5XFAD Alzheimer Mice at Different Disease Stages. Neuroscience, 307, 128-137.

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